 
 
 
Risk Management Proposal  
 
Bovine Germplasm   
BOVIGERM.GEN  
 
 
 
 
[Document Date]   
  
 
 
Disclaimer  
This risk management proposal does not constitute, and should not be regarded as, legal advice. While every 
effort has been made to ensure the information in this document is accurate, the Ministry for Primary Industries 
does not accept any responsibility or liability whatsoever for any error of fact, omission, interpretation or opinion 
that may be p resent, however it may have occurred.  
© Crown Copyright - Ministry for Primary Industries  
 
 
 
 
 
 
 
 
 
 
 
 
 
Contact for further information : 
Ministry for Primary Industries (MPI)  
Biosecurity New Zealand  
Animal Imports  
PO Box 2526  
Wellington 6140  
Email: animal.imports@mpi.govt.nz  
Telephone:  04 890 0134  
 Risk Management Proposal:  Bovine Germplasm  [Document Date ] 
Ministry for Primary Industries   Page 1 Contents  
1 Purpose  3 
2 Background  3 
3 Objective  4 
4 General requirements for all importations of bovine germplasm  5 
4.1 Application  5 
4.2 Diagnostic tests, vaccines and treatments  5 
4.3 Semen collection centre requirements  5 
4.4 Donor requirements  6 
4.5 Semen collection, processing and storage  6 
4.6 Embryo collection, processing and storage  7 
4.7 Transport  8 
5 Recommendations for identified risk organisms  8 
5.1 Bluetongue virus (BT)  8 
5.2 Borna disease  9 
5.3 Bovine herpes virus 1.1, 1.2a, and 5 (infectious bovine rhinotracheitis/infectious pustular 
vulvovaginitis, IBR/IPV)  9 
5.4 Bovine leukaemia virus, BLV (enzootic bovine leukosis, EBL)  10 
5.5 Bovine viral diarrhoea virus genotype 2 (BVDV2)  12 
5.6 Crimean Congo haemorrhagic fever virus (CCHF)  14 
5.7 Foot and mouth disease virus (FMD)  15 
5.8 Lumpy skin disease virus (LSD)  16 
5.9 Rift Valley fever virus (RVF)  17 
5.10  Vesicular stomatitis virus (VS)  17 
5.11  Brucella melit ensis, Brucella suis, and Brucella abortus (bovine brucellosis)  18 
5.12  Campylobacter fetus  subspecies venerealis  (bovine genital ca mpylobacteriosis, BGC)  19 
5.13  Chlamydophila abortus  20 
5.14  Coxiella burnetii  (Q-fever)  21 
5.15  Leptospira  interrogans serovar hardjoprajitno (leptospirosis)  22 
5.16  Mycobacterium tuberculosis  (bovine tuberculosis)  23 
5.17  Mycoplasma mycoides  subsp. mycoides  SC (contagious bovine pleuropneumonia, CBPP) 24 
5.18  Other mollicutes ( Mycoplasma  and Ureaplasma  spp.)  24 
5.19  Exotic Salmonella  spp. (salmonellosis)  29 
6 Appendice s 30 
6.1 Appendix 1: Comparative Efficacy Study  30 
6.2 Appendix 2: Antibiotic Treatment of Bovine Semen  33 
 
  
Ministry for Primary Industries   Page 3 1 Purpose  
The purpose of this document is to:  
• Show how options for the management of risk o rganisms have been considered . 
• Provide recommendations for import requirements.  
2 Background  
Bovine semen and embryos have the potential to harbour exotic viral and bacterial diseases. In February 2009, 
the Ministry of Agriculture and Forestry (now the Ministry for Primary Industries, (MPI) completed the Import Risk 
Analysis (IRA) Cat tle Germplasm from All Countries . This import risk analysis (2009 IRA) forms the basis of the 
risk management measures in the proposed import health standards (IHSs) for bovine semen and embryos.  
Following on from that IRA, the Import  Health Standard: Bovine Semen, BOVSEMID.GE N and Import Health 
Standard: Bovine Embryos , BOVEMID.GE N were published on 27 June 2011.  
These IHSs contain generic import requirements which manage the biosecurity risk asso ciated with importing 
bovine semen and in vivo derived bovine embryos from MPI approved countries.  
It is proposed that the existing IHSs are combined into one IHS and the proposed title and short -code are Semen 
and In Vivo Embryos from Bovines, BOVIGERM.G EN. The guidance document (GD) will be updated by MPI prior 
to issuing the IHS. It provides commodity -specific guidance information including links to country -specific 
bilaterally -agreed veterinary certification for trade in bovine semen and in vivo derive d bovine embryos.  
Generic IHSs served as the basis for country to country (bilateral) negotiations of country -specific veterinary 
certificates. MPI agreed country specific veterinary certificates with the exporting country’s Competent Authority 
once MPI wa s satisfied with the exporting country’s export systems. Negotiations took into account the verifiable 
health status of the exporting country, the national systems, legislation and processes in the exporting country for 
regulatory oversight of the bovine g ermplasm industry, and the capabilities and preferences of the exporting 
country’s Competent Authority. The assessments were based on the World Organisation for Animal Health OIE 
Code (the OIE Code ) section 3, Quality of Veterinary Services . 
Bovine semen c ountry -specific veterinary certificates are currently available for Australia, USA, Canada, 
European Union, Norway and Switzerland. Bovine embryo country specific veterinary certificates are currently 
available for Australia, USA, Canada, the European Unio n and Switzerland.  
Where the IHS requires that the OIE Code  recommendations are met, the requirements reflect the current OIE 
Code . When OIE Code  chapters are amended, MPI reviews these changes to ensure they continue to align with 
New Zealand’s appropriat e level of protection. Where OIE Code  recommendations no longer meet New 
Zealand’s appropriate level of protection, they are replaced with risk -based MPI recommendations and the IHSs 
will be amended. Otherwise the most recent version of the OIE Code  is ref erred to.  
Since the first IRA in 2009, new scientific information and changes to New Zealand’s disease status have 
prompted new risk assessments. The outcomes of those assessments are discussed in this RMP for 
incorporation into the new IHS. Those assessme nts include . 
• Rapid Risk Assessment:  Mycoplasma bovis in bovine semen  in 2019  (RRA bovine semen 2019).   
• Rapid Risk Assessment: Mycoplasma bovis in bovine in -vivo and in -vitro produced embryos  (RRA  bovine 
embry os 2018).   
• Import Risk Analysis: Bovine Leukaemia Vir us and  Campylobacter fetus  Subspecies venerealis  in Bovine 
Frozen Semen, In -vivo and In -Vitro Produced Embryos  (IRA BLV and C. fetus  2018).  
• Rapid Risk Assessment: Vesicular Stomatitis Virus in Live Animals and Their Germplasm .   
Ministry for Primary Industries   Page 4 Additionally, technical advice was provided from MPI’s Risk Analysis team on bovine viral diarrhoea virus 
(BVDV2), and Crimean Congo haemorrhagic fever (CCHF).  
This RMP discusses ways to incorporate the new risk advice into the e xisting IHS. Due to the ongoing 
Mycoplasma bovis  eradication programme, there was a preliminary consultation on the  Discussion Document : 
Mycoplasma  bovis risk management in bovine germplasm  from 23 September to 5 November 2019. This RMP 
describes all proposed amendments in the IHS, including  those for  M. bovis .   
The IHS was initially consulted May -July 2020 and subsequently provisionally released on 30 September 2020. 
At that time new scien ce was published that was relevant to the risk management of M. bovis  in semen. MPI has 
subsequently reviewed this evidence and sought an expert review and is now undertaking a second consultation 
of the revised IHS.  
Expert advice was received from Dr Paul  Chambers (veterinary pharmacologist, Massey University) in February 
2021 regarding antibiotic treatment of semen for M. bovis control. It is attached to this document in Section 6, 
Appendix 2.   
3 Objective  
The objective is to manage, to an appropriate level  of protection, the biosecurity risks for bovine semen and 
bovine embryos, consistent with New Zealand’s domestic legislation and international obligations.  
New Zealand has adhered to the Application of Sanitary and Phytosanitary Measures (the SPS Agreemen t) for 
over two decades, and in doing so recognises the desirability of using agreed international standards for trade 
wherever possible. Under Article 2.1 of the World Organisation for Animal Health (OIE) Code  Chapter on Import 
Risk Analysis, “The objecti ve is to manage risk appropriately to ensure that a balance is achieved between a 
country’s desire to minimise the likelihood or frequency of disease incursions and their consequences and its 
desire to import commodities and fulfil its obligation under int ernational trade agreements.” And under Article 5.3  
of the SPS Agreement , “Members may choose to implement sanitary measures more stringent than those in 
international standards, if these are deemed necessary to protect animal or human health and are scien tifically 
justified by a risk analysis.”    
Of the hazards identified in the import risk analyses, the 2011 IHS included risk management for the following 
hazards:  
• Bluetongue virus  
• Borna disease virus  
• Exotic bovine herpes viruses (semen only)  
• Bovine viral  diarrhoea virus type 2  
• Crimean Congo haemorrhagic fever  
• Foot and mouth disease virus  
• Lumpy skin disease virus  
• Rift Valley fever virus  
• Vesicular stomatitis virus  
• Exotic Brucella  spp. (semen only)  
• Coxiella burnetii  
• Exotic Leptospira  spp.  
• Mycobacterium bov is 
• Mycoplasma mycoides  subsp. mycoides  SC 
• Other exotic Mycoplasma  spp.  
Risk management was not considered justified in 2011 for the following hazards identified by the IRA:  
• Campylobacter fetus  subspecies venerealis  (semen only)  
• Chlamydophila abortus   
Ministry for Primary Industries   Page 5 • Bovine leukaemia virus (semen only)  
• Exotic Salmonella  spp. 
Identified hazards proposed for addition to the IHS in the 2020 amendment include:  
• Bovine leukaemia virus  
• Campylobacter fetus subspecies venerealis   
Identified hazards proposed for removal in the 2020 amendment include:  
• Bluetongue virus  
• Borna disease vir us 
• Crimean Congo haemorrhagic fever  
• Vesicular stomatitis virus  
4 General requirements for all importations of bovine 
germplasm  
The general requirements section of the IHSs include risk management measures for semen and embryo donors, 
regardless of the countr y’s disease freedom claims. Where possible, these requirements align with the 
recommendations of the OIE Code  and the International Embryo Transfer Society (IETS) Manual.  
4.1 Application  
(1) The IHS applies to frozen semen and in vivo derived  embryos from the Bovi nae subfamily for import from 
all countries into New Zealand.  
4.2 Diagnostic tests, vaccines and treatments  
(1) All pre -export and/or surveillance testing required by the IHS must be:  
a) conducted by a laboratory approved by the Competent Authority of the exporting  country; or  
b) conducted by a laboratory approved by the Competent Authority of any other country approved to 
export the specified type of bovine germplasm  to New Zealand.  
(2) All laboratory samples required by the IHS must be collected, processed, and stored in  accordance with 
the recommendations in the OIE Code  and/or the Manual  or as described in the document, Approved 
Diagnostic Tests, Vaccines, Treatments and Post -arrival Testing Laboratories for Animal Import Health 
Standards, MPI-STD-TVTL .   
(3) All diagnostic test(s) and vaccines that are required to be used or undertaken by the IHS must be those 
that have been approved by MPI for that purpose and documented in MPI-STD-TVTL .  
(4) All products and vaccinations required by the IHS to be administered to meet the specific disease 
requirements in Part 2 of the IHS must have been administered according to the manufacturer’s instruction  
in a c ountry that the CTO has agreed meets the requirements of clause 1.5  of the IHS.  
(5) All requirements in th e IHS for the administration of a vaccine require that either the final dose of a primary 
vaccination course or the recommended booster to complement the primary course  is administered . 
4.3 Semen c ollection centre requirements  
(1) Semen collection must be carried out in a semen collection centre that complies with the 
recommendations for centres in the Code  chapter General Hygiene in Semen Collection  and Processing 
Centres.   
Ministry for Primary Industries   Page 6 (2) The semen collection centre must be:  
a) approved for export by the exporting country’s Competent Authority;  
b) subjected to regular inspection, at least every 12 months, by an Official Veterinarian;  
c) under the supervision of a semen coll ection centre veterinarian .  
(3) Semen donors may be transferred from one approved semen collection centre to another approved centre 
of equal health status without isolation or testing if the Competent Authority of the exporting country 
ensures that all the f ollowing requirements are met:  
a) Donors are examined by the approved semen collection centre veterinarian on the day of entry into 
the centre and show no evidence of infectious disease transmissible in semen.  
b) Transfer is direct.  
c) Donors do not come into direc t or indirect contact with animals of lower health status.  
d) The means of transport is disinfected before use.  
4.4 Donor requirements  
(1) Semen donors must meet the requirements in the Code  chapter Collection and Processing of Bovine, 
Small Ruminant, and Porcine Semen , and any additional requirements in Part 2  of the IHS. 
(2) During the 28 days in which semen donors are held in pre -entry isolation prior to entering the semen 
collection centre (as prescribed in the OIE Code ), they must not be used for natural m ating, and must be 
isolated from animals not of equivalent health status.  
(3) Embryo donors must meet the recommendations in the Code  chapter Collection and Processing of In Vivo 
Derived Embryos from Livestock and Equids  and any additional requirements in Part 2 of the IHS. 
(4) Embryo donors must be resident in the embryo collection herd for at least 28 days prior to embryo 
collection for export to New Zealand  until completion of pre-collection donor testing . While resident with 
the collection herd, the herd must not be subject to veterinary restrictions for the identified risk organisms 
managed in Part 2 of the IHS.  
(5) Germplasm d onors that were imported to the exporting country must have lived continuously in an 
approved country for at least 60 days before germplas m collection.  
(6) On the day of germplasm collection, the embryo collection team veterinarian or semen collection centre 
veterinarian must determine that the donor is free from clinical evidence of infectious diseases 
transmissible in germplasm.  
(7) Where a speci fic requirement of the IHS for a risk organism is met by pre -collection testing, the 
germplasm donors must be isolated from other animals not of equivalent tested health status, from the 
time of the pre -collection test until completion of collection for ex port.  
(8) Where a specific requirement of the IHS for a risk organism is met by monitoring the germplasm donor for 
clinical signs for a specified time after collection, the germplasm must be stored for that amount of time 
prior to export.  
4.5 Semen collection, processing and storage  
(1) Semen collection, processing and storage must comply with the sections relevant for bovine semen in the 
Code  chapter Collection and Processing of Bovine, Small Ruminant, and Porcine Semen , unless 
otherwise stated in Part 2 of this IH S. 
(2) Where Part 2 of the IHS requires testing within a certain time period before or after semen collection:  
a) Semen collection may be  a time period of up to 60 consecutive days.  
b) Samples for testing before collection must be obtained within the specified period before the first day 
of the semen collection period.  
c) Tests required after semen collection must have samples collected within the specified period after 
the last day of the semen collection period.   
Ministry for Primary Industries   Page 7 (3) A cryogenic or cooling agent used in the  freezing process, storage and transport must not have been used 
previously in association with any other product of animal origin.  
(4) All straws must be sealed, and clearly and permanently marked to identify the donor and the date(s) of 
freezing. The markin gs must conform to international standards of the International Committee for Animal 
Recording (ICAR) . If a code is used for this information, its decipher instructions must accompany the 
consignment.  
(5) Semen must  only be stored  and transported  with germplas m that has been collected and processed in 
accordance with the Code .  
(6) Semen must be held in a storage place approved by the Competent Authority until the time of export.  
(7) Subject to (8), semen may only be imported into New Zealand if the semen is imported directly from the 
country in which it was collected.  
(8) If semen is collected in a country that meets the requirements of 1.5 of this IHS and stored in another 
country that also meets the requirements of 1.5 of this IHS (country of storage), that semen may b e 
imported into New Zealand if the consignment is accompanied by:  
a) A declaration from the Competent Authority of the country of storage that:  
i) identifies the semen from the country of origin as the semen being exported to New Zealand;  
ii) certifies that the se men has been stored and transported in the country of storage in 
accordance with the requirements of this IHS.  
b) Evidence that the semen meets the rest of the requirements of this IHS in the form of either:  
i) a veterinary certificate issued by the Competent Authority of the origin country; or  
ii) a letter from the Competent Authority of the origin country confirming the semen meets the 
requirements of this IHS and indicating which requirements therein have been fulfilled.    
4.6 Embryo collection, processing and stor age 
(1) Embryos must be collected, washed, processed, stored and traceability maintained under the supervision 
of an embryo collection team veterinarian and in accordance with:  
a) the recommendations in the OIE Code  chapter on Collection and Processing of In Vivo Derived 
Embryos from Livestock and Equids ; 
b) the recommendations in the IETS Manual . 
(2) The embryo collection team must operate in accordance with the conditions listed in the  OIE Code  
chapters on Collection and Processing o f In Vivo Derived Embryos from Livestock and Equids .   
(3) At the time of embryo collection each embryo must be examined over its entire surface at not less than 
50X magnification and found to have an intact zona pellu cida and be free of adherent material.  
(4) Any micro -manipulation that causes a breach of the zona pellucida  must be done as per the procedures 
described in the OIE Code  chapter Collection and Processing of Micromanipulated Oocytes or Embryos 
from Livestock and Horses  and the IETS Manual .  
(5) All biological products of animal origin used in the media and solutions for collection, processing, washing 
or storage of embryos must be free of pathogenic organisms including pestiviruses and prions. Media and 
solutions must be sterilised by approved me thods according to the IETS Manual  and handled in a manner 
that ensure s sterility is maintained.  
(6) All straws must be sealed, and clearly and permanently marked to identify the donor and the date(s) of 
freezing. The markings must conform to international st andards of the International Committee for Animal 
Recording  (ICAR; www.icar.org) and the IETS  Manual . If a code is used for this information, its decipher 
instructions must accompany the consignment.  
(7) Embryos must  only be stored and transported with germpl asm that has been collected and processed in 
accordance with the OIE Code .   
Ministry for Primary Industries   Page 8 (8) Embryos must only be held in a storage place approved by the Competent Authority until the time of 
export.  
(9) Subject to (1 0), embryos can only be imported into New Zealand if the em bryos are imported directly from 
the country in which they were collected.  
(10) If embryos are collected in a country that meets the requirements of 1.5 of this IHS and stored in another 
country that also meets the requirements of 1.5 of this IHS (country of storage), those embryos may be 
imported into New Zealand if the consignment i s accompanied by:  
a) A declaration from the Competent Authority of the country of storage that:  
i) identifies the embryos from the country of origin as the embryos being exported to New 
Zealand;  
ii) certifies that the embryos have been stored and transported in th e country of storage in 
accordance with the requirements of this HIS.  
b) Evidence that the embryos meet the rest of the requirements of this IHS in the form of either:  
i) a veterinary certificate issued by the Competent Authority of the origin; or  
ii) a letter fr om the Competent Authority of the origin country confirming the embryos meet the 
requirements of this IHS and indicating which requirements therein have been fulfilled.   
4.7 Transport  
(1) All transport containers for transport to New Zealand must be new or disinf ected, and confirmed empty 
prior to loading. When a transport container is disinfected, the disinfectant, its active chemical and the date 
of disinfection must be recorded on the veterinary certificate.  
(2) All transport containers in which germplasm is transp orted to New Zealand must be sealed, by either the 
collection centre veterinarian or an Official Veterinarian, using tamper -evident seals that are positioned to 
ensure that no germplasm can be added after the transport container  has been sealed. The seal n umber 
must be recorded on the veterinary certificate.   
(3) Where germplasm is transferred from one transport container to another, the date of transfer, approved 
collection centre /herd  or storage facility , reason for transfer, and the name of veterinarian inv olved in the 
transfer must be recorded on the veterinary certificate.   
5 Recommendations for identified risk organisms  
5.1 Bluetongue virus (BT) 
5.1.1 Risk management options presented in the 2009 IRA  
The 2009 IRA assessed  the risk of bluetongue virus for bovine germp lasm to be  negligible.  
5.1.2 Discussion  
(1) Measures have been maintained to align with international recommendations. However , measures are no 
longer considered justified. Several factors led to this conclusion:  
a) New Zealand is free of Culicoides  vectors, the only known vectors capable of transmitting BTV.  
b) New Zealand monitors for Culicoides , including monitoring sentinel cattle for seroconversion to 
arboviruses.  
c) According to the OIE Code , New Zealand will not lose its bluetongue free status thr ough the 
importation of vaccinated, seropositive or infective animals, or semen or embryos/ova from infected 
countries or infected zones.  
d) The vir aemic period in cattle can be up to 11 weeks. However , cattle are not carriers.    
e) Bluetongue is not a zoonotic  disease   
Ministry for Primary Industries   Page 9 5.1.3 Recommendation  
(1) Measures are not justified.  
5.2 Borna disease  
5.2.1 Risk management options presented in the 2009 IRA  
(1) Require germplasm donors to have been resident since birth in countries where the disease has never 
been reported.   
(2) Require donors to origi nate from herds with a greater than 5 year history of freedom from the disease in 
countries in which the disease is notifiable or in which reliable histories are available.  
(3) Test aliquots of semen and embryos from each collection by intracerebral inoculatio n into rabbits or by 
culture on cell cultures from embryonic rabbit or rat brain with negative results.  
(4) PCR test of peripheral cells from donors with negative results ( accuracy and reliability of this testing 
methodology have been questioned.)  
(5) It is propos ed that all of the options suggested in the risk analysis be used, in modified form, in the import 
health standard.   
5.2.2 Discussion  
(1) Borna disease (BD) is not an OIE listed disease.  
(2) There is no evidence that it is venereally transmitted in any species of anim al and the public health impact 
remains only speculative. Measures were excluded from the IHSs for ovine and equine germplasm, and 
those are the species primarily affected by the disease.    
5.2.3 Recommendation  
(1) Measures are not justified.  
5.3 Bovine herpes virus 1. 1, 1.2a, and 5 ( infectious bovine 
rhinotracheitis/ infectious pustular vulvovaginitis, IBR/IPV)  
5.3.1 Risk management options presented in the 2009 IRA  
(1) Test each batch of semen by a VI test (or a suitable PCR test when available).  
(2) Require that donors be from herds or breeding centres that are maintained free from IBR in accordance 
with the OIE Code . 
(3) Require isolation of donor animals for at least 30 days after semen collection and testing by a validated 
serological test with negative result, at least 21 days after semen collection.  
5.3.2 Discussion  
(1) Abortions and encephalitis have not been associated with BHV in New Zealand, where only BHV1.2b has 
been isolated. Both syndromes of BHV infection, IBR and IPV, are present in New Zealand .    
(2) Chronically infected latent carriers of BHV exist and are known to excrete BHV in their semen.  
(3) A 2005 study demonstrated that seronegative breeding bulls can produce BHV infected semen. There 
appears to be little other research that confirms this fin ding. It is generally agreed that the OIE Code  
recommendations appropriately manage the risk in bovine semen originating from AI centres that meet the 
OIE Code’s  hygiene, collection, and processing recommendations.   
Ministry for Primary Industries   Page 10 (4) After decades of importing under the exi sting measures, the IHS should continue to align with the OIE 
Code . Aligning with the Code  requires that the 30 -day post -collection isolation period is added to the IHS’s 
post-collection test requirement.  
(5) The virus is known to adhere to embryos, but it is  removed when IETS washing protocol (with trypsin) is 
followed. It is an IETS category 1 disease and embryos should be collected, processed and stored in 
accordance with the OIE Code . No additional donor or germplasm testing is recommended.  
(6) BHV5 is poorly understood, although it h as been detected  in bovine semen and is believed to cause 
disease, including vulvovaginitis . Its diagnosis is not certain using the approved tests  for BHV1.1 and 1.2a . 
BHV5 is rare and  characterised by neurological signs that begin  about 10 days post -infection ; however, 
the infection can remain latent.  Most serological tests for BHV 1.1 and 1.2a are believed to be capable of 
detect ing BHV5 antibodies as well.  
(7) Due to the severity of BHV5  clinical signs, the stress bulls experience moving  from the ir herd of origin to a 
semen collection centre  [stress is believed  to be a trigger for the neurological signs to develop] , veterinary 
oversight of collection centres, the potential for transmission within a  herd, and the short incubation period, 
it is considered  that the disease would be observed at the collection centre, during the period before 
collection. The risk can therefore be managed via historical collection centre freedom.   
5.3.3 Recommendation  
(1) At the time of collection of semen for export to New Zealand, the exporting country must be free from BHV 
1.1 and BHV 1.2a in accordance with the OIE Code ; or 
(2) The semen collection centre must be maintained free from BHV 1.1 and 1.2a from commencement until 
conclusion of semen colle ction for export to New Zealand, through compliance with the recommendations 
in the OIE Code  in relation to BHV, with the following modifications:  
a) Test all cattle prior to pre -entry isolation for antibodies using a test listed in MPI-STD-TVTL , with 
negative results;   
b) Test all cattle in pre -entry isolation for antibodies using a test listed in MPI-STD-TVTL , with negative 
results, or where an animal in a group has tested positive re -testing the remaining animals, with 
negative results, not less than 21 days after removal of the positive anim al;  
c) Thereafter, annually re -test all cattle  for antibodies using a test listed in MPI-STD-TVTL , with 
negative results; or  
(3) The semen donor must be:  
a) held in isolation for the 30 days following colle ction;  
b) tested for BHV 1.1 and 1.2a using a test listed in MPI-STD-TVTL  at least 21 days after semen 
collection for export to New Zealand, with negative results; or  
(4) An aliquot of semen from each sem en collection for export to New Zealand must be tested for BHV 1.1 
and 1.2a using a test listed in MPI-STD-TVTL , with negative results. . 
(5) The following should be require d for BHV  5:  
a) The semen donor’s centres in residence have had no cases of BHV  5 (suspected or diagnosed) in 
the year  prior to collection for export to New Zealand .     
(6) Measures are not justified for embryos.  
5.4 Bovine leukaemia virus, BLV ( enzootic bovine leukosis, EBL)  
5.4.1 Risk m anagement options presented in the 2018 IRA  
(1) The semen was collected, processed and stored in accordance with Chapters 4.6 and 4.7 of the OIE 
Code . 
Note: This is a general requirement for all imported semen.   
Ministry for Primary Industries   Page 11 (2) The semen was collected, processed and stored in  accordance with Chapters 4.6 and 4.7 of the OIE 
Code ; and  
a) An aliquot of not less than 0.5 ml of processed semen from the final collection of each donor was 
tested by an approved virus isolation test or a polymerase chain reaction test with negative result s. 
 
Note: This option would likely further manage the risk of infected semen over and above that achieved by 
Option 1 alone.  
(3) The semen was collected, processed  and stored in accordance with Chapters 4.6 and 4.7 of the OIE 
Code ; and  
a) The donor bull was resident at the time of semen collection in an Enzootic Bovine Leukosis (EBL) 
free herd in accordance with the OIE Code . 
b) If less than two years of age, the bull cam e from a serologically negative ‘uterine’ dam; or  
i) The bull was subjected to diagnostic tests for EBL on blood samples on two occasions with 
negative results, the first test being carried out at least 30 days before and the second test at 
least 90 days afte r collection of the semen.  
Note: This option aligns with the OIE Code  recommendation for the importation of bovine semen.  
5.4.2 Discussion  
(1) The preliminary hazard list comp iled in the 2009 IRA considered bovine leukaemia virus as endemic in 
New Zealand, therefore , no risk management was recommended (2009 IRA).   
(2) In 1997 New Zealand dairy industry implemented the “Dairy Enzootic Bovine Leukosis” scheme. Under 
this scheme the last BLV -infected cows were culled in 2007 -2008. Since then there have been no 
detections of BLV in New Zealand and annual screening of the da iry industry was maintained at over 50% 
of all herds for several years (Voges, 2012)1. Export testing of cattle, including beef breeds,  over several 
years  has produced no positive results.  
(3) BLV, the causative agent of EBL is notifiable under the Biosecuri ty (Notifiable Organisms) Order 2016.  
(4) BLV is subject to passive surveillance in New Zealand.  
(5) Transmission of BLV occurs through the transfer of infected lymphocytes and BLV proviral DNA has been 
isolated in semen.  Venereal transmission by natural service and artificial insemination has not been 
demonstrated.  
(6) According to Import Risk Analysis: Bovine Leukaemia Virus and Campylobacter fetus Subspecies 
venerealis in Bovine Frozen Semen, In -vivo and In -Vitro Produced Embryos (IRA BLV and C. fetus 2018) , 
the likelihood of BLV being present in semen  is low. The likelihood of subsequent exposure and 
transmission of BLV to susceptible animals is assessed to be low. The consequences of entry and 
establishment of BLV are assessed to be non -negligible. Bovine leukaemia virus is therefore assessed to 
be a risk in imported bovine semen. Risk management is therefore justified.  
(7) Seroconversion can take up to 3 months after infection.  
(8) The OIE Code  provides recommendations for the importation of bovine semen. New Zealand has 
achieved and maintained freedom in t he absence of risk management. The risk management 
recommended by the Code  (two tests over 4 months  and herd freedom ) is considered out of proportion 
with the risk .  
(9) Examination  of the seminal vesicles should be considered a useful strategy for the detecti on of 
inflammation that would contribute lymphocytes to semen.  If herd freedom is established, the risk should 
be considered managed.       
(10) A PCR on semen has been developed and is considered to be more sensitive than virus isolation.  
 
1 Voges, (2012). Reports from industry surveillance and disease control programmes: New Zealand Dairy Enzootic Bovine Leukosis (EBL) Control Scheme, 
Surveillance, 39 (3):41   
Ministry for Primary Industries   Page 12 (11) If only semen were tes ted, a sample from every collection would need to be tested. Testing diluted semen 
is considered acceptable due to low risk of BLV being present in semen.  
(12) The 2018 IRA concluded that BLV is not a risk in in vivo derived bovine embryos. The washing protocols 
have been shown to be effective to remove embryo -associated BLV (IETS Category 1).  
(13) The IHS should not require measures for countries that a CTO recognises as free from EBL.  
5.4.3 Recommendation  
(1) At the time of semen  collection for export to New Zealand, the exporting country must be recognised by a 
CTO as free from EBL; or  
(2) The semen donor must be resident at the time of semen collection in a herd certified as free from EBL by 
the Competent Authority; and  
a) If less than  two years of age, the semen donor must come from a ‘uterine dam’ that has been 
subjected to serological test listed in MPI-STD-TVTL ; or 
b) The semen donor must originate from a herd certified as free by the Competent Authority; or  
c) The semen donor must be subjected to a test listed in MPI-STD-TVTL  for EBL, with negative results, 
between 30 days before and 30 days after collection of the semen; or  
(3) The semen donor must be subjected to a test listed in MPI-STD-TVTL  for EBL on two occasions with 
negative results, the first test being carried out at least 30 days before and the second test at le ast 90 
days after collection of the semen; or   
(4) The semen donor must be subjected to a test listed in MPI-STD-TVTL  for EBL, with negative results, 
between 30 days before and 30 days after collection  of the semen; and  
a) the semen donor must pass a breeding soundness exam, including manual rectal palpation or visible 
observation with rectal ultrasound of the seminal vesicles, conducted on the day of collection by a 
collection centre veterinarian; or  
(5) An aliquot of semen from each collection for export to New Zealand must be tested for BLV using a test 
listed in MPI-STD-TVTL , with negative results.   
(6) Note : there are no import requirements for embryos . 
5.5 Bovine viral diarrhoea virus genotype 2 (BVDV2)  
5.5.1 Risk management options presented in the 2009 IRA  
(1) Require donor bulls to have been resident on a semen collection centre where bulls are maintained and 
tested as specified in the OIE Code  chapter on collection and processing of bovine semen. [This includes 
testing the collection herd prior to pre -entry is olation by virus isolation or virus antigen test and a 
serological test, then full repeat testing in pre -entry isolation].  
(2) Require each batch of semen be tested by VI or reverse transcriptase polymerase chain reaction (RT -
PCR).   
(3) Require that embryo donors  be tested as bulls are in semen collection centres (as per OIE Code  chapter 
on collection and processing of bovine semen).  
(4) Require non -fertilized, degenerated, and zona pellucida compromised embryos, collection fluid and 
washing fluid (or an embryo from t he first embryo collection for each consignment) from each donor be 
collected and be tested by VI or RT -PCR, with negative result.   
5.5.2 Discussion  
(1) BVDV2 has not been described in cattle in New Zealand.  
(2) Both acutely infected and persistently infected semen do nors need to be prevented from supplying semen 
for export to New Zealand.   
Ministry for Primary Industries   Page 13 (3) The antigen detection tests are not appropriate for detecting acutely infected cattle due to their transient 
viraemia. Semen donors should be tested for antibodies at least 3 weeks apart (prior to pre -entry isolation 
and in pre -entry isolation) to detect acutely infected donors. During isolation, acutely infected cattle would 
be expected to show a 4 -fold rise in antibody titre.   
(4) If any semen donor is serologically positive prior to e ntering isolation  in a BVDV2 free herd , the donor 
must have returned a negative semen collection while resident in the collection centre.  
(5) BVDV is an IETS Category 3 disease2 with regard to  the risk of disease transmission via in vivo  derived 
embryos. A 2013 study showed that in vivo  produced embryos infected with BVDV when exposed to 
BVDV infected semen do not transmit the virus to recipients or offspring when washed with trypsin 
according to IETS washing requirements. Embryos should be considered lower risk than semen.  
(6) A 21 day isolation period for embryo donors should be considered to cover the incubation period (3 -7 
days) for acute BVD infections. As BVDV -2 can cause severe disease, anim als displaying clinical signs 
will be identified and excluded as embryo donors. Acutely infected animals would be expected to clear the 
virus prior to collection.   
(7) Persistently infected cattle do not have detectable antibodies, therefore the antigen detec tion ELISA, real -
time RT -PCR and virus isolation, as described in the OIE Manual , should be required. Semen donors 
should be tested twice, 3 -4 weeks apart.  
(8) Embryo donors need only be subjected to a single viral or antigen test at any time  to be confident that they 
are not persistently infected. The donor should be isolated from untested cattle between the test date and 
embryo collection  or tested for antibody to confirm the donor was not acutely infected at the time of 
collection . 
(9) Animals which remain in c ollection centres should be tested annually.    
(10) Germplasm can be tested by PCR. Test results from a pooled embryo sample or semen sample from the 
same day of collection can be considered representative of the collection.   
(11) For semen , it should be clarified  that testing should be within the 30 days prior to the start of pre -entry 
isolation. It should also be clarified that seropositive donors which have been in a free herd at a collection 
centre need only have a single negative test on semen while resident i n the centre. The way in which 
BVDV can invade embryos and resist washing, indicates a need for the semen used to fertilise embryos 
for export to meet the semen requirements of the IHS.  
(12) Vaccination has been shown to cause a transient viraemia for up to 10  days. It is unknown if or how the 
virus may be shed after vaccination. A precautionary measure to manage the transient viraemia (waiting 
30 days after vaccination before collection) is warranted.  
(13) If post collection testing is necessary , serology is consi dered to offer reliable sensitivity . The test can be 
conducted several months after collection because titres can be detected for years .  
(14) It is recognised that in rare cases, BVDV has been demonstrated to sequester in the genital tract. In these 
cases, det ection is only possible via germplasm testing. In these rare cases, it is believed that the  donor’s 
germplasm would fail industry quality standards .  
5.5.3 Recommendation  
(1) At the time of germplasm collection for export to New Zealand, the exporting country must b e recognised 
by a CTO as free from BVDV2; or  
(2) Semen:  
 
2 According t o the IETS Manual, Category 3 diseases or pathogenic agents are those for which preliminary evidence indicates that the risk of transmission is 
negligible provided that the embryos are properly handled between collection and transfer in accordance with the  IETS Manual, but for which additional in 
vitro and in vivo experimental data are required to substantiate the preliminary findings.   
Ministry for Primary Industries   Page 14 a) The semen collection centre must be maintained free from BVDV2 from commencement until 
conclusion of semen collection for export to New Zealand, in accordance with the recommendations 
in the OIE Code  in relation to BVDV, with the following modifications:   
i) Test all cattle within 30 days prior to pre -entry isolation using a test listed in MPI-STD-TVTL ;  
ii) Test all cattle after 21 days isolation using a test listed in MPI-STD-TVTL ; 
iii) If any animal seroconverts, keep all animals in pre -entry isolation until there is no more 
seroconversion for 3 weeks;   
iv) Only approve entry for groups where pre -entry isolation results indicate the absence of 
antigen -positive cattle;   
v) Annually re -test seronegative cattle using a test listed in MPI-STD-TVTL ;  
vi) For seropositive donors, test semen for BVDV 2 using a test listed in MPI-STD-TVTL  with 
negative results, prior to collection for export to New Zealand; or  
b) An aliquot of semen from each semen collection for export to New Zealand must be tested for 
BVDV2 using a test lis ted in MPI-STD-TVTL , with negative results.  
(3) Embryos:  
a) A sample of the unfiltered collection fluid or an embryo from the collection for export to New Zealand 
must be tested for BVDV2 in accordance wit h MPI-STD-TVTL  with negative results; or  
b) The embryo donor must be tested for persistent BVDV2 infection using a test listed in MPI-STD-
TVTL , with negative results; and the  
i) semen used to produce the embryo must satisfy the requirements of this IHS;  
ii) embryo donor must not have been vaccinated against BVDV2 in the last 30 days;  
iii) embryo donor must be tested for acute BVDV2 infection in accordance  with MPI-STD-TVTL , 
with negative results, in one of the following ways:  
1) using the antigen capture ELISA test immediately prior to an isolation period of at least 
21 days before collection for New Zealand. Isolation must exclude cattle that were not 
tested negative for BVDV2 upon entry to the collection herd, and throughout isolation 
the herd showed no clinical signs consistent with BVDV2; or  
2) using the virus isolation test within 48 hours of collec tion for New Zealand; or  
3) serologically tested between 2 weeks and 6 months after collection.      
5.6 Crimean Congo haemorrhagic fever virus (CCHF)  
5.6.1 Risk management options presented in the 2009 IRA  
(1) Require germplasm donors to have been resident for at least the 21 days before germplasm collection in a 
country or zone that is free from the disease.  
(2) Require scrupulous treatment of germplasm donors with acaricide, and inspection and placement in tick 
free germplasm collection premises. Donors would be required t o be quarantined for at least 3 weeks prior 
to start off, and during germplasm collection with regular inspection and maintenance of tick free status.  
(3) Require serological testing of germplasm donors within 7 days prior to start of germplasm collection and 3-
8 weeks after germplasm collection.   
5.6.2 Discussion  
(1) Presence of the virus or transmission of the virus by insemination or implantation of germplasm (of any 
species) has never been described.  
(2) Hyalomma ticks are considered to be required for the establishment of the disease. There is no evidence 
that the New  Zealand cattle tick is a competent vector.   
(3) Although never previously demonstrated, if a recipient dam were to become viraemic with CCHF virus, 
she would remain so for only up to 7 days. There are no descriptions of long term carriers. During 
viraemia, h er blood would pose a zoonotic risk.   
Ministry for Primary Industries   Page 15 (4) Technical advice received in 2016 concluded that ruminant germplasm is not a viable pathway for CCHFV 
to enter New Zealand.  
5.6.3 Recommendation  
(1) Measures are not justified.  
5.7 Foot and mouth disease virus (FMD)  
5.7.1 Risk managemen t options presented in the 2009 IRA  
(1) Institute measures in accordance with the OIE Code  for semen and embryos.  
(2) Prohibit importation of germplasm from countries that are infected with foot and mouth disease.  
5.7.2 Discussion  
(1) Due to the extreme seriousness of the  disease and the catastrophic consequences that would result from 
its introduction, it was concluded that importation of bovine germplasm should be limited to countries or 
zones that are free of FMD virus in accordance with the OIE Code , or countries or zo nes in which 
compliance with measures in accordance with the recommendations of the OIE Code  for import of 
germplasm from FMD infected countries or zones has been reviewed and accepted by MPI.  
(2) The OIE Code’s  recommendation for importing from a FMD free cou ntry include monitoring the semen 
donor for 30 days after collection for clinical signs of FMD. Therefore , all semen must be frozen for 30 
days before export to New Zealand.  
(3) While the OIE Code  indicates that no additional measures are justified for embryo s, the measures 
recommended for semen from free countries or zones are:  
(4) Semen Donors:  
a) showed no clinical signs of FMD on the day of collection of the semen and for the following 30 days;  
b) were kept in a centre where no animal had been added in the 30 days  before collection, and that 
FMD has not occurred within a 10-kilometre  radius of the centre for the 30 days before and after 
collection; either  
i) have been vaccinated at least twice, with the last vaccination not less than one month and not 
more than six mo nths prior to collection, unless protective immunity has been demonstrated 
for more than six months; or  
ii) were subjected, not less than 21 days after collection of the semen, to tests for antibodies 
against FMDV, with negative results;  
(5) Semen:  
a) was collected, processed and stored in accordance with Chapters 4.6 and 4.7 of the Code ;  
b) was subjected, with negative results, to a test for evidence of FMDV if the donor males has  been 
vaccinated within the 12 months prior to collection;  
c) was stored in the country of origin for a period of at least one month following collection, and that 
during this period no animal on the establishment where the donor males were kept showed any sign 
of FMD.  
5.7.3 Recomm endation  
(1) No change in risk management from the current IHS’s FMD requirements.  
(2) Semen:  
a) The donor must be resident for at least the 3 months before the semen collection in a country or 
zone that is free from FMD without vaccination in accordance with the OIE Code ; or  
Ministry for Primary Industries   Page 16 b) The herd of origin, semen collection centre, donor animal and semen for export to New Zealand 
must comply with OIE Code  recommendations for export of bovine semen from countries or zones 
presenting a risk of FMD; and  
i) Each semen collection, processing and storage facility in the exporting country intended to be 
used during the preparation of an export consignment to New Zealand must be approved by a 
CTO.  
(3) Embryos:  
a) The donor must be resident for at least the 3 months before e mbryo collection in a country or zone 
that is free from FMD without vaccination in accordance with the OIE Code ; or  
b) The herd of origin, the embryo collection herd where the donors were resident during embryo 
collection, the donor animal and the embryos fo r export to New Zealand must comply with the OIE 
Code  FMD Article Recommendations for the Importation of In Vivo Derived Embryos of Cattle ; and  
i) Each embryo collection, processing and storage facility in the exporting country, intended to 
be used during the  preparation of an export consignment to New Zealand, must be approved 
by a CTO.  
Note: The approval will be dependent on the establishment, its location and operating standards, and that 
the verification systems of the veterinary authority achieve a very h igh level of risk management for FMD. 
The process for MPI approval may include site inspection. MPI reserves the right to supervise collection or 
require any other measures deemed necessary to ensure compliance with facility and operating standards 
upon wh ich the approval is based.  
5.8 Lumpy skin disease virus (LSD)  
5.8.1 Risk management options presented in the 2009 IRA  
(1) Donors have been resident for 6 months prior to germplasm collection in a country or zone that is free of 
LSD as defined by the OIE Code . 
(2) Donors c ould be required to be resident in an establishment or germplasm collection centre that has been 
free from LSD for at least 6 months. All animals on the centre could be required to be free from any sign of 
LSD for at least 28 days after completion of germp lasm collection.  
(3) Aliquots of semen and embryo wash fluid, substandard embryos or aliquot of embryos, from each batch of 
imported germplasm could be tested for LSD .  
5.8.2 Discussion  
(1) Each of the options presented in the import risk analysis were included, in modified form, in the 
recommended options for the IHS.  
5.8.3 Recommendation  
(1) No change from t he current IHS LSD requirements.  
(2) The germplasm donor must be resident for 6 months prior to germplasm collection in a country or zone 
that is free of LSD as defined by the OIE Code ; or 
(3) The germplasm donor must be resident in an establishment that was free of clinical evidence of LSD 
during a period from at least 6 months prior to commencement, until 28 days after conclusion of 
germplasm collection for export t o New Zealand; or  
(4) An aliquot of semen or a sample of embryos/oocytes, collection fluids and/or washing fluids from each 
germplasm collection for export to New Zealand must be tested for LSD using  a test listed in MPI-STD-
TVTL  with negative results.   
Ministry for Primary Industries   Page 17 5.9 Rift Valley fever virus (RVF)  
5.9.1  Risk management options presented in the 2009 IRA  
(1) Donors have been resident for 30 days prior to collection of germplasm, and during germplasm collection, 
in an RVF free co untry or zone.  
(2) Donors have been resident for 6 months prior to and during collection of germplasm in a n RVF infected 
country in which climatic changes predisposing to RVF outbreaks have not occurred in the previous 6 
months.  
(3) Donors were held in mosquito -free premises at least 30 days prior to, and during collection.  
5.9.2 Discussion  
(1) The country freedom option presented in the import risk analysis was included in the IHS. The other 
options in the IHS were worded to align more closely with the OIE Code . The Code  RVF chapter has been 
amended since 2011 and the new recommendations should be adopted. They now allow for donors to be 
seropositive or vaccinated. The risk is considered to be managed since long term carriers of the virus have 
not been described. Vaccination  would need to be in accordance with the OIE.      
5.9.3 Recommendation  
(1) The donor must be resident, for at least the 30 days prior to, and during germplasm collection for export to 
New Zealand in a country or zone that is free from RVF in accordance with the OIE  Code ; or 
(2) The donor must not show any clinical signs of RVF within the period from 14 days prior to and 14 days 
following germplasm collection; and either  
a) the donor must be vaccinated against RVF at least 14 days prior to collection; or  
b) the donor must be demonstrated to be seropositive on the day of collection using a test listed in MPI-
STD-TVTL ; or 
c) testing of paired samples using a test listed in MPI-STD-TVTL  must demonstrate that 
seroconversion did not occur between germplasm collection and 14 days after.  
5.10 Vesicular stomatitis virus (VS)  
5.10.1  Risk estimation presented in the 2016 rapid risk assessment:  
(1) For animal germplasm, the likelihood of entry is assessed to be negligible. Accordingly, the risk estimate 
for animal germplasm is negligible.  
5.10.2  Discussion  
(1) OIE has removed VS from its notifiable disease list.  
(2) There is a considerable body of evidence showing that viraemia does not occur in VS, introduction via 
germplasm is therefore not assessed to be  a risk . There is also no carrier status.  
(3) Additionally, the infection is primarily insec t-borne and there are no known vectors present in New 
Zealand.   
5.10.3   Recommendation  
(1) It is recommended the measures for VS are removed in alignment with the OIE Code . No specific 
measures are necessary.   
Ministry for Primary Industries   Page 18 5.11 Brucella melitensis , Brucella suis, and Brucella abortus (bovine 
brucellosis)  
5.11.1   Risk management options presented in the 2009 IRA  
(1) Donor bulls could be required to be kept since birth in a country or zone that is officially free from 
brucellosis.  
(2) Donor bulls could be housed at an artificial breeding centre where  the testing programme for bulls includes 
testing with both the complement fixation test and the buffered antigen agglutination test.  
(3) Donor bulls could be required to be kept in a herd officially free from bovine brucellosis, showed no clinical 
sign of bov ine brucellosis on the day of collection of the semen and were subjected to a buffered Brucella 
antigen test with negative results during the 30 days prior to collection. ( Note : This option is for bulls not 
maintained on an approved semen collection centre . The standard will require that bulls are housed on a 
semen collection centre.)  
(4) Donor bulls were kept in a herd free from bovine brucellosis, showed no clinical sign of bovine brucellosis 
on the day of collection and were subjected to the buffered Brucell a antigen and complement fixation tests 
with negative results during the 30 days prior to collection.  ( Note : This option is for bulls not maintained 
on an approved semen collection centre.)  
(5) Each of the options suggested in the import risk analysis were in cluded as recommended options in the 
import health standard in modified form.   
5.11.2   Discussion  
(1) According to the OIE (2019) Brucella melitensis and Brucella suis have never been reported in New 
Zealand. Brucella abortus is known to be absent in New Zealand, an d its last occurrence in domestic and 
wild animal population was recorded in 1989.  
(2) Brucella abortus, Brucella suis,  and Brucella melitensis,  causative agents of bovine brucellosis, are 
unwanted, notifiable organisms in New Zealand.  
(3) The second option in th e IRA should be  modified to require maintenance of semen collection centre 
freedom from brucellosis in accordance with the recommendations of the OIE Code  chapter on collection 
and processing of bovine semen.   
(4) There is evidence of B. suis infection in cat tle and although there is no evidence of the organism in bovine 
semen, the risk is considered non -negligible because Brucella spp . have an affinity for the reproductive 
tract.  
(5) Although the OIE Code  Chapter on bovine brucellosis has incorporated B. suis, vertical and horizontal 
transmission of B. suis  in cattle is rare and risk management should reflect the degree of risk.   
(6) A single test for seroconversion after collection is an appropriate alternative to herd freedom  for B. suis .  
(7) Bovine brucellosis  is not a hazard in bovine embryos.  
5.11.3  Recommendation  
(1) The B. melitensis and B. abortus risk should be managed with the following options:  
a) The semen donor must be kept since birth in a country or zone that is free from Brucella in 
accordance with the OIE Code ; or 
b) The semen collection centre must be maintained free from Brucella from commencement until 
conclusion of semen collection for export  to New Zealand, through compliance with the 
recommendations in the OIE Code  in relation to Brucella , with the following modifications:  
i) The donor must be from a country , zone , or herd  that is free from Brucella in accordance with 
the OIE Code  or the Compet ent Authority;  
ii) During the 30 days prior to pre -entry isolation, the donor must be tested using a test listed in 
MPI-STD-TVTL  for Brucella , with negative results;   
Ministry for Primary Industries   Page 19 iii) All cattle in pre -entry isolation must be tested using a test listed in MPI-STD-TVTL  for Brucella , 
with negative results;  
iv) At least annually all cattle resident in the semen collection centre must be  tested using a test 
listed in MPI-STD-TVTL  for Brucella , with negative results; or  
c) The semen collection centre must be maintained free from Brucella from commencement until 
conclusion of semen coll ection for export to New Zealand, through compliance with the Competent 
Authority’s collection centre testing program in relation to Brucella . 
(2) The Brucella  suis risk should be managed with the following options:  
a) The semen donor must be kept since birth in a country or zone that is recognised by a CTO as free 
from Brucella suis ; or 
b) The semen collection centre must be maintained free from Brucella suis  from commencement until 
conclusion of semen collection for export to New Zealand, through compliance with th e Competent 
Authority’s collection centre testing program in relation to Brucella suis ; or 
c) The donor must be tested using a test listed in MPI-STD-TVTL  for Brucella suis  between 2 weeks 
after entry into the collection centre and 90 days after collection, with negative results.  
5.12 Campylobacter fetus  subspecies venerealis  (bovine genital 
campylobacteriosis, BGC)  
5.12.1  Risk management options presented in the 2018 risk work  
(1) Veterinary Authorities of importing countries should require the presentation of an international veterinary 
certificate attesting that the donor animals:  
a) Have never been used for natural service; or  
b) Have only mated virgin heifers; or  
c) Were kept in an artificial insemination centre where no c ase of bovine genital campylobacteriosis 
(BGC) has been reported; and  
d) The culture of semen and preputial specimens for the presence of the causal agent of bovine genital 
campylobacteriosis proved negative.  
 
Note: This option aligns with the recommendations provided by OIE Code . Some of the included sub 
options may be difficult to certify. The option should only be considered where BGC is notifiable.  
(2) The semen was collected, processed and stored in accordance  with Chapters 4.6 and 4.7 of the OIE 
Code ; and  
a) The donor animal has been kept in a pre -entry isolation facility for at least 28 days and after a 
minimum of 7 days in pre -entry isolation has undergone testing for Campylobacter fetus  subspecies 
venerealis  as follows:  
b) Animals less than six months old or kept since that age only in a single sex group prior to pre -entry 
isolation should be tested once on a preputial specimen, with a negative result.  
c) Animals aged six months or older that could have h ad contact with females prior to pre -entry 
isolation should be tested three times at weekly intervals on a preputial specimen, with a negative 
result in each case.  
d) All resident bulls and teasers in the semen collection facility are tested at least annually  for 
Campylobacter fetus subspecies venerealis , with negative results, where the country or zone where 
the semen collection facilities are located is not free;  
i) A preputial specimen should be tested.  
ii) Only bulls on semen production or having contact with bul ls on semen production need to be 
tested. Bulls returning to collection after a lay -off of more than six months should be tested not 
more than 30 days prior to resuming production.  
 
Note:  This aligns with the recommendations for the importation of bovine s emen provided by OIE 
Code  Chapter 4.6.   
Ministry for Primary Industries   Page 20 5.12.2   Discussion  
(1) Campylobacter fetus  subspecies venerealis , which is the causative organism of bovine genital 
campylobacteriosis (BCG), was endemic in New Zealand and the 2009 IRA did not recommend risk 
mitigation.  
(2) There have been no laboratory confirmed isolations of Campylobacter fetus  subspecies venerealis  in New 
Zealand since 1992 (Loveridge & Gardner 1993)3. 
(3) Campylobacter fetus  subspecies venerealis  is notifiable in New Zealand under the Biosecurity (Notifiable 
Organ isms) Order 2016.  
(4) The IRA 2018 assessed  that the likelihood of entry of Campylobacter fetus  subspecies venerealis  in in 
vivo derived bovine embryos, processed as per the IETS protocols is negligible. Accordingly,  the IRA 2018 
concluded that risk management  measures for Campylobacter fetus  subspecies venerealis  in imported 
bovine embryos are not required. The OIE Code  also does not recommend any measures for bovine 
embryos.  
(5) BGC is an OIE listed disease (OIE 2018) and the OIE Code  chapter 4.6 recommends test ing bulls and 
teaser animals used for producing semen for Campylobacter fetus  subspecies venerealis . Chapter 11.3, 
Article 4, provides specific recommendations for bovine semen imports.  
(6) The IRA 2018 assessed Campylobacter fetus  subspecies venerealis  in se men to be a risk and risk 
management is scientifically justified.  
(7) In recommending risk management options for Campylobacter fetus  subspecies venerealis  in bovine 
semen, the IRA 2018 noted  that: 
a) Campylobacter fetus subspecies venerealis  is transmitted thro ugh natural service and AI using 
infected bull semen.  
b) Infection in bulls is not associated with clinical signs, gross lesions, histological changes or altered 
semen quality.  
c) In 2011 susceptibility to antibiotics generally used in semen treatment was limite d to gentamicin.  
(8) If exporting countries have legislation and/or BCG control programmes in place at collection centres 
approved for export, those should be  able to be considered for approval.  
(9) Risk management via vaccination is not recommended at this time because they are considered to only 
provide short -lived immunity and  may not terminate an established infection.      
5.12.3  Recommendation  
(1) At the time of collection for New Zealand, the semen collection centre must have a programme, approved 
by the Competen t Authority or aligned with the OIE Code , to prevent Cfv infected animals from entering 
the collection herd, and there must have been no case of bovine genital campylobacteriosis reported in 
centre in the 1 year prior to  collection; or  
(2) A preputial or semen  sample from the donor must be tested  for Cvf using a listed in MPI-STD-TVTL  in the 
last 6 months, and all tests conducted in the last 6 months must have been negative.  
5.13 Chlamydophila abortus  
5.13.1   Risk  management options presented in the 2009 IRA  
(1) Donors resident since birth or for the previous 2 years in a country or zone free from Chlamydophila 
abortus  based on no laboratory confirmation of infection in any species for at least 2 years.  
(2) Individual dono rs could be tested serologically using an OIE recommended test for C. abortus , 2-3 weeks 
after germplasm collection.  
 
3 Loveridge, R., Gardner, E., (1993) Campylobacter fetus venerealis  infection in cattle, Surveillance Surveillance Vol.20 No.4.   
Ministry for Primary Industries   Page 21 (3) Aliquots of germplasm could be tested for C. abortus  by PCR or antigen detection ELISA, with negative 
results.  
(4) Tetracycline or macrolide an tibiotics could be added to imported germplasm.  
5.13.2   Discussion  
(1) The literature on C. abortus  infection in cattle does not provide evidence of transmission from cattle to 
other susceptible animal species or humans. The literature indicates that C. abortus  has been associated 
with abortion in cattle, but transmission appears to be limited to periods of close association between 
cattle and infected sheep or goats, or where bedding from animals infected with C. abortus  is recycled in 
dairy herd housing.  
(2) In one st udy it was shown that infected ewes did not produce embryos that led to infection of recipients or 
progeny. There have  not been studies to assess transmission from infected bovine embryos, but it is 
considered unlikely and there are no documented cases of transmission.      
(3) Experimental studies have shown that C. abortus  can be excreted in semen of inoculated bulls and 
isolation of the agent from semen of naturally infected bulls and rams has been reported. Transmission 
from semen has not been demonstrated.     
(4) While sheep are considered long term carriers of C. abortus , it is unknown if cattle can be long term 
carriers of infection and if they would harbour the organism in the reproductive tract.  
(5) All human cases of abortion caused by C. abortus  have been as sociated with direct contact with small 
ruminants.  
(6) Establishment in New Zealand would have significant impacts on sheep farming and human health.  
(7) It was concluded in 2010 that measures specifically against Chlamydophila  should not be placed in the 
import  health standard for bovine germplasm.  
5.13.3   Recommendation  
(1) Measures are not justified.  
5.14 Coxiella burnetii  (Q-fever)  
5.14.1  Risk management options presented in the 2009 IRA  
(1) Require acaricide treatment and subsequent inspections to demonstrate tick freedom during the 30-day 
quarantine of germplasm donors.  
(2) Donors could be tested by an ELISA test, with negative results, 21 -60 days after the final collection of 
germplasm for exp ort to New Zealand.  Consider prohibiting germplasm from any animals that have ever 
been known to test positive.  
(3) MPI approved test on each collection of germplasm.  
5.14.2   Discussion  
(1) Acaricide treatment and 30-day quarantine in tick free premises would add consid erable cost to the import 
of bovine germplasm.  
(2) Serological testing of donors has been used historically to mitigate risk. The IDEXX ELISA test has been 
validated for cattle and shown to provide acceptable sensitivity4; other ELISA tests will require MPI 
approval during veterinary certificate negotiation.  
 
4 R Kittelberger , J Mars , G Wibberley , R Sting , K Henning , GW Horner , KM Garnett , MJ Hannah , JA Jenner , CJ Pigott & JS  O'Keefe (2009) 
Comparison of the Qfever complement fixation test an d two commercial enzyme -linked immunosorbent assays for the detection of serum antibodies against 
Coxiellaburnetii (Q -fever) in ruminants: Recommendations for use of serological tests on imported animals in New Zealand, New Zealand Veterinary 
Journal, 57:5 , 262 -268, DOI: 10.1080/00480169.2009.58619   
Ministry for Primary Industries   Page 22 (3) There was a 2016 study indicating that the indirect fluorescence antibody (IFA) assay may be a superior 
option for testing ruminants.  
(4) The IHS should be written to allow for MPI approval of other test met hods in the future.  
(5) An option for whole herd testing for Q fever was incorporated in the 2011 IHS. This option is useful for 
countries conducting herd monitoring on a routine basis.    
5.14.3   Recommendation  
(1) The germplasm donor must never have been confirmed pos itive for Q fever; and either  
a) The donor must be subjected to a serological test listed in MPI-STD-TVTL  for Q fever, on a sample 
collected between 21 and 120 days after each germplasm collection for export to New Zealand, with 
negative results; or  
b) An aliquot of semen or a sample of embryos/oocytes, collection fluids and/or washing fluids from 
each germplasm collection for export to New Zealand must be tested for Q fever using a test listed in 
MPI-STD-TVTL , with negative results; or  
c) Within the 6 -month period before or after germplasm collection for New Zealand, but before export, 
the embryo collection herd or semen collection centre herd must be tested for Q fever, using a test 
listed in MPI-STD-TVTL , with negative results. The Q fever test must be:  
i) performed on either the whole herd or a random sample of at least 60 animals (whichever is 
the lesser number); and  
ii) The herd must be isolated for the period between semen collection and diagnostic sampling.  
5.15 Leptospira  interrogans serovar hardjoprajitno  (leptospirosis)  
5.15.1   Risk management options presented in the 2009 IRA  
(1) Serological testing of don ors to demonstrate freedom from exotic Leptospira  spp. 
(2) PCR or culture of germplasm for exotic Leptospira  spp. 
(3) Preparation of germplasm in accordance with the recommendations of the OIE Code , including the use of 
suitable antibiotics in semen diluents and embryo washing media.  
5.15.2   Discussion  
(1) Despite extensive surveys, there have been no detections of Leptospira  interrogans serovar hardjoprajitno  
in New Zealand.  
(2) Serovar hardjoprajitno is transmitted venereally  and has been associated with abortion storms in cattle. 
Cattle are most likely the maintenance hosts for this serovar.  
(3) Leptospira spp. are commonly excreted in the semen of bulls and are known to adhere and penetrate 
embryos after in vitro exposure.  
(4) Leptospira  spp. are sensitive to a variety of antibiotics . Inclusion of antibiotics in prepared semen has 
traditionally been used to prevent dissemination of Leptospira  spp. by international trade. Antibiotic 
treatment of embryos is likely to be effective.  
(5) For semen the antibiotics recommended as per the Code  (at the doses recommended or higher)  would 
manage the risk.   
(6) For embryos the antibiotic combinations indicated in the IETS Manual would manage the risk.  
(7) Serological testing, specifically the microscopi c agglutination test (MAT), could be considered for risk 
management depending on its ability to detect chronic carriers.   
5.15.3   Recommendation  
(1) For semen minimum doses of antibiotics as per the OIE Code  Article 4.7.7 .    
Ministry for Primary Industries   Page 23 (2) For embryos  minimum doses of antibiotics as per the IETS Manual :  
a) 100 IU penicillin and 100 μg streptomycin; or  
b) 100 IU penicillin and 50 μg gentamicin; or 
c) 200 µg tylosin  
(3) Alternative  antibiotic  combinations effective against Leptospira spp. could be considered  for addition to 
MPI-STD-TVTL  during vet erinary certificate negotiation.  
(4) Serological test ing could be considered for addition to  MPI-STD-TVTL  during  veterinary certificate 
negotiation.   
5.16 Mycobacterium tuberculosis  (bovine tuberculosis)  
5.16.1   Risk management options presented in the 2009 IRA  
(1) Semen donors could be required to show no clinical sign of bovine tuberculosis on the day of collection of 
the seme n. 
(2) Semen donors could be required to be kept in an artificial insemination centre free from bovine 
tuberculosis in a country, zone or compartment free from bovine tuberculosis and which only accepts 
animals from free herds in a free country, zone or compar tment as defined by the OIE.  
(3) Semen donors could be required to show negative results to tuberculin tests carried out annually and be 
kept in a herd free from bovine tuberculosis as defined by the OIE.  
(4) Embryo donors and all other susceptible animals in the herd of origin could be required to show no clinical 
sign of bovine tuberculosis during the 24 hours prior to embryo collection.  
(5) Embryo donors could be required to have originated from a herd free from bovine tuberculosis in a 
country, zone or compartm ent free from bovine tuberculosis.  
(6) Embryo donors could be required to be kept in a herd free from bovine tuberculosis, be isolated in the 
establishment of origin for the 30 days prior to departure to the collection centre and be subjected to a 
tuberculin t est for bovine tuberculosis with negative results.  
5.16.2   Discussion  
(1) In accordance with suggested options in the import risk analysis, alignment with the OIE Code  and the 
New Zealand bovine tuberculosis pest management strategy was chosen for management of the r isk 
associated with bovine tuberculosis for bovine germplasm imported to New Zealand.  
5.16.3   Recommendation  
(1) Semen:  
a) At the time of collection for New Zealand, the semen collection centre must be:  
i) free from bovine tuberculosis in accordance with the OIE Code  or the Competent Authority of 
the exporting country;  
ii) located in a country or zone that was recognised by a CTO as free from bovine tuberculosis . 
b) The semen collection centre must be maintained free from bovine tuberculosis from commencement 
until conclusion of semen collection for export to New Zealand, in accordance with the 
recommendations in the OIE Code  in relation to bovine tuberculosis;  
i) Prior  to pre -entry isolation, donors must be from a herd free from bovine tuberculosis, either in 
accordance with the OIE Code  or the Competent Authority of the exporting country;  
ii) At least annually all resident cattle must be tested using a test listed in MPI-STD-TVTL  for 
bovine tuberculosis, with negative results.  
(2) Embryos:   
Ministry for Primary Industries   Page 24 a) There must not have been any clinical signs of bovine tuberculosis observed in the embryo collection 
herd during the 24 hours prior t o embryo collection for export to New Zealand;  
b) The donor must be from an embryo collection herd that was free from bovine tuberculosis at the time 
of collection for export to New Zealand in accordance with the OIE Code  or the Competent Authority 
of the ex porting country; and the donor must be : 
i) from a country or zone recognised by a CTO as free from bovine tuberculosis; or  
ii) subjected to a test listed in MPI-STD-TVTL  for bovine tuberculosis during the period between 
30 days prior to and 12 months after embryo collection for export to New Zealand, with 
negative results.  
5.17 Mycoplasma mycoides  subsp. mycoides  SC (contagious bovine 
pleuropneumonia, CBPP)  
5.17.1   Risk management options presented in the 2009 IRA   
(1) Donors of germplasm could be required to originate from a country or zone that is free from CBPP.  
(2) Donors could be required to not be vaccinated against CBPP and be kept since birth or for at least 6 
months in an establishment where no case of CBPP has been  reported and the establishment is not 
situated in a CBPP infected zone.  
(3) Donors could be subjected to an OIE recommended serological test with negative results on two 
occasions 21 -30 days apart with the last test done within 14 days prior to germplasm coll ection.  
5.17.2   Discussion  
(1) CBPP is caused by Mycoplasma mycoides  subsp. mycoides  SC 
(2) For the purposes of the OIE Code , the incubation period is 6 months. Studies have shown the incubation 
period to be between 3 weeks and 3 months.   
(3) Each of the options presented i n the import risk analysis were included, in modified form, in the 
recommended options for the 2011 IHS. The second and third options were included as a combination of 
requirements for countries infected with CBPP.  
5.17.3   Recommendation  
(1) The germplasm donor must be born in and have been continuously resident in a country that is recognised 
by a CTO as free from CBPP; or  
(2) The germplasm donor must:  
a) never have been vaccinated for CBPP;  
b) be kept since birth, or for at least the 6 months prior to commencement until concl usion of 
germplasm collection for export to New Zealand in establishments where no case of CBPP has been 
reported, and which are not situated in a CBPP infected zone, as defined by the OIE Code ; 
c) be serologically tested for CBPP, using a test listed in MPI-STD-TVTL  on two occasions 21 to 30 
days apart, with the last test within 14 days prior to germplasm collection for export to New Zealand, 
with negative results.  
5.18 Other mollicutes ( Mycoplasma  and Ureaplasma  spp.)  
5.18.1   Risk management options presented in the 2009 IRA   
(1) The 2009 IRA recommended the following options for management of Mollicutes ( Mycoplasma 
bovigenitalium, Mycoplasma bovis, Mycoplasma verecundum, Mycoplasma californicum, Mycoplasma 
canaden se, Mycoplasma group 7  and Ureplasma diversum ). 
a) Addition of an appropriate combination of antibiotics to semen extenders.   
Ministry for Primary Industries   Page 25 b) Culture of germplasm for identification of organisms before adding antibiotics.  
c) Culture of germplasm for identification of organisms a fter addition of antibiotics.  
5.18.2   Discussion  
Ureaplasma  spp.  
(1) Ureaplasmas are present in New Zealand, however it is unknown which species are present.  
(2) Measures are not considered justified.  
Mycoplasma bovis risk management since 2011  
(3) The OIE Code  does not have a chapter on Mycoplasma  spp. and there is no reporting requirement.  
(4) The current M. bovis  requirement was incorporated in 2011. It indicates that the donor has never recorded 
a positive test for M. bovis.  It does not require M. bovis  testing . MPI has considered the M. bovis  risk 
managed via the requirement that donors (and the semen collection centres or collection herd) had to 
meet the general health requirements as set out by the OIE Code  and semen and embryos had to be 
collected and proces sed in accordance with the OIE Code , including specifications for the addition of 
antibiotics.  
(5) The OIE Code’s  recommended antibiotic combinations are (antibiotic dose is per mL frozen semen):  
a) Gentamicin (250 µg), tylosin (50 µg), lincomycin –spectinomycin (150/300 µg) (GTLS);  
b) Penicillin (500 IU), streptomycin (500 µg), lincomycin -spectinomycin (150/300 µg);  
c) Amikacin (75 µg), divekacin (25 µg).  
(6) The OIE Code  does not recommend a specific protocol for the use of antibiotics during semen or embryo 
processing.  
(7) The OIE Code  references the International Embryo Technology Society (IETS) Manual  for embryo 
processing and antimicrobial use. An effective approach to managing M. bovis  contamination is discussed 
in the IETS Manual , but it is not specifically recommended by IETS and would not have been carried out 
on exported embryos under the current terms of trade since targeted antibiotic treatment is not explicitly 
stated in the IHS.   
New Zealand’s  M. bovis status  
(8) Genetic analy sis of recovered M. bovis  strains provide evidence that M. bovis  was not endemic prior to the 
initial detection in 2017 and all isolates sequenced to date are consistent with  a single strain being  
introduced into New Zealand in 2015/16.  
(9) Surveillance data shows that most new infected properties are identified through tracing, rather than 
general surveillance, signalling that M. bovis has not become endemic and that multiple geographically 
widespread entry events or systematic failure had not occurred. Surve illance data supports the findings of 
the genetic analysis.  
(10) The eradication programme strategy therefore focuses on:  
a) tracing movements from affected properties to new farms  
b) identifying additional infected properties through a National Surveillance Programme . 
(11) Disease spread in New Zealand is managed by movement controls on properties that are at risk or 
infected. Moveme nt controls stay in place until either:  
a) testing shows that the herd is not infected  
b) infected properties are depopulated, then cleaned and disinfected.  
(12) Germplasm collection centres have not been among the infected properties.  
(13) M. bovis risk management in bovine germplasm is warranted due to New Zealand’s commitment to 
eradication, it’s non-endemic status , and the non -negligible risk.    
Ministry for Primary Industries   Page 26 Discussion Document: Mycoplasma bovis Risk Management in Bovine Germplasm  
(14) The consultation on the  document: Discussion Document: Mycoplasma bovis Risk Management in Bovine 
Germplasm  ended on 5 November 2019 . It:  
a) explains the eradication p rogramme and why MPI attributes New Zealand’s M. bovis freedom until 
2017, after decades of imports of over 2.5 million straws, to be due to the low risk of trans mission 
and use of antibiotics;  
b) communicates that it is unknown how M. bovis  entered New Zeala nd; 
c) discusses semen processing and antibiotic treatment, as well as other risk mitigation strategies such 
as testing.  
(15) The submissions were considered during the development of the M. bovis measures in the draft import 
health standard, consulted on in May 2 020.  
Testing for  M. bovis  
(16) Experimentally, M. bovis  antibodies can be detected in serum by indirect ELISA 6 to 10 days after 
infection. However, under natural conditions, individual animal titres are not well correlated with infection 
or disease, meaning antibodies may not be detected in infected animals. Moreover, the ELISA test is 
validated as a herd detection tool rather than for use in individual animals and its estimated diagnostic 
sensitiv ity is around 75% .   
(17) Low diagnostic sensitivity of the availab le tests and the persistent subclinical  infection d oes not permit 
screening of donors by serological tests to confirm freedom from M. bovis infection.   
(18) The major challenge to current molecular detection tests is the intermittent shedding of M. bovis  and t he 
absence of validated tests . PCR results on samples cannot be considered a reliable indicator of donor 
freedom from infection with M. bovis  for the purpose of international trade . 
(19) The Discussion Document: Mycoplasma bovis Risk Management in Bovine Germpl asm mentions a 
validated PCR test for semen . Such a test would allow individual batches  of semen that test negative to be  
deemed safe for trade. However , validation is incomplete, and the sensitivity of the PCR test  on semen is 
unknown.  
(20) MPI is undertaking laboratory validation of the PCR using M. bovis spiked semen.  
(21) A reverse transcriptase  PCR (RT-PCR) targeting RNA to focus detection methods on viable  M. bovis  may 
be longer term solution. Validation work would be required.      
Current semen a ntibiotic treatment methods  
(22) The USA’s Certified Semen Services (CSS) claims that their antibiotic protocol s, CSS Minimum 
Requirements , are effective against M. bovis . They  reference a 1988 study conducted on inoculated 
semen.  
(23) The antibiotic  options recommended by the OIE (including the GTLS doses) are not supported by 
literature as effective for M. bovis  control.  
(24) In Appendix  1 of the CSS Minimum  Requirements  provides two methods for antibiotic treatment. Both 
methods require neat/raw semen treatment with 500 µg gentamicin, 100 µg tylosin, 300/600 µg Linco -
Spectin (GTLS) for 3 -5 minutes before the addition of any extender/glycerol. Both methods al so require 
that semen is held in contact with the antibiotics for 2 hours above 5°C. MPI considers that the neat 
semen treatment (both the dose and duration), the time before freezing, and freezing process are the 
critical elements of their methods to achi eve effective management of M. bovis . These elements are 
common to both CSS methods.  
May 2020 IHS consultation  
(25) The draft IHS required the GTLS combination of antibiotics at double the OIE dose (per mL 
extended/frozen semen), aligning with the CSS’s 1-step method, as well as measures to ensure contact 
time with the antibiotics prior to freezing.    
Ministry for Primary Industries   Page 27 (26) A submitter provided another study (see Appendix 1) comparing the two CSS treatment methods. The two 
methods were shown to produce similar M. bovis control despite  different final doses. Both methods use 
the same protocol  on neat semen treatment.   
(27) The submissions highlighted the lack of literature to support an antibiotic requirement focusing on the final 
concentration of antibiotics in extended semen. MPI subsequently found evidence that the extenders 
interfere with antibiotic efficacy. MPI has not  found evidence to support the withholding glycerol in the 2 -
step method.   
(28) Both methods provide the best documented control of M. bovis .  
(29) As discussed in Discussion Document: Mycoplasma bovis Risk Management in Bovine Germplasm , 
several studies have shown that M. bovis is developing greater antimicrobial resistance. MPI’s 
communications with M. bovis experts has revealed the perception that CSS’s methods may not be 
appropriate for future risk management.  
(30) MPI has noted that the study in Appendix 1 shows the  bactericidal effect of CSS’ 2 -step method 
decreased from approximately 70%  (in 1988)  to approximately 53% in 2000 . The  efficacy of the GTLS 
combination in 2000 was less than 100%.  
September 2020 p rovisional IHS  
(31) MPI issued an amended, provisional IHS after considering all submissions  and the study from 2000 .  
(32) MPI proposed  to empirically i ncreas e the antibiotic dose  (by 20%)  and the incubation period (from a 
minimum of 3 minutes to 5 minutes) for neat semen treatmen t due to evidence of declining efficacy more 
than two decades ago combined with evidence of growing resistance. The point of maximum efficacy had 
not been evaluated in the previous studies.  
(33) Treating neat/raw semen is considered to be a more targeted appro ach that is expected to provide better 
bactericidal effect  and better overall antibacterial control . It allows the exporter more flexibility in terms of 
how the semen is extended , it uses smaller overall volume compared to requiring a higher final dose  and 
was not expected to significantly affect sperm viability  based on what had been reported in the literature.  
(34) For embryos, MPI decided to adhere to the protocol described in the IETS Manual.    
(35) During the provisional period  a new study funded by the Minist ry of Agriculture and Forestry of Finland5 
was published .    
March 2021 IHS consultation  
(36) The new study looked specifically at the efficacy of the GTLS antibiotics on M. bovis in semen. MPI 
decided to review the study and undertake another round of consult ation .   
(37) MPI agreed with the new study ’s conclusion tha t it is challenging to rely on the use of antibiotics in bovine 
semen production to control M. bovis . The amount of natural M. bovis contamination in semen is largely 
unknown, but t he amount of contamination is critical to the control that is likely to be achieved under the 
different treatment methods. The new study showed that e ven at low contamination the CSS GTLS dose 
for neat semen cannot be considered to provide an appropriate l evel of protection for New Zealand.  
(38) MPI commissioned a veterinary pharmacologist, Dr Paul Chambers, to provide expert opinion on the 
empirical antibiotic treatment (see Appendix 2 of this document). Dr Chamber’s opinion is that a dose 
higher than CSS’s do se is likely to likely to inhibit a greater proportion of the bacteria present. He also 
explained the dose and time dependent elements of the GTLS combination.  
(39) Until there is a validated test for M. bovis in semen  or for donor bulls , a higher dose and lon ger incubation 
should  more effectively manage the increasing risk (due to M. bovis resistance) .  
(40) An exporter reported to MPI that an initial t rial of the proposed  20% increase d antibiotic dose was proving 
to have a significant impact o n post-thaw semen motility.  
 
5 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7601584/   
Ministry for Primary Industries   Page 28 (41) In response , MPI has readjusted the dose to 15% over the CSS dose and decreased the neat semen 
incubation time from 5 minutes to 4 minutes. CSS requires a 3 -5 minute neat semen incubation.   
(42) MPI’s Animal Health Laboratory is developing a study that  will investigate alternative treatment protocols 
for bovine semen contaminated with M. bovis.  
(43) The embryo requirements are unchanged from those indicated in the IETS Manual as appropriate for 
treating M. bovis  as MPI are not aware of any other studies that demonstrate effective management in 
embryos.   
5.18.3   Recommendation  
(1) Semen:  
a) Collection and processing of the semen must be in accordance with the recommendations of the OIE 
Code , with the following modifications:  
i) The raw/neat semen for export to New  Zealand must have one of the following combinations 
added to it at the specified dose per mL of neat/raw semen:  
1) gentamicin ( 575 µg), tylosin (1 15 µg), lincomycin –spectinomycin (3 45/690 µg) (GTLS); 
or 
2) another MPI approved antibiotic combination, listed in MPI-STD-TVTL ;  
ii) The antibiotics must be either:  
1) Prepared and stored as separate stock solutions as described by the manufacturer to 
maintain pote ncy; or  
2) Premixed and used as indicated by the manufacturer to retain potency;  
iii) The semen and antibiotic solution must not be further diluted for at least 4 minutes;  
iv) The semen must remain at no less than 5°C for a minimum of 2 hours before being frozen in 
the antibiotic solution; or  
b) The semen must be subjected to another MPI approved method of processing, listed in MPI-STD-
TVTL ; or 
c) The semen donor must be subjected to a test for M. bovis listed in MPI-STD-TVTL , with negative 
results; or  
d) Each semen collection for export to New Zealand must be tested using a validated PCR test for M. 
bovis  listed in MPI-STD-TVTL , with negative results.  
 
(2) Embryos:  
a) Collection and processing of the embryos must be in accordance with the recommendations of the 
OIE Code , with the following modification:  
i) The embryos must be subjected to the protocol described in the IETS Manual : tylosin (200 
µg/mL) incubation at 37°C in the antibiotic treatment for a minimum of 4 hours after being 
washed 10 times; or  
b) The embryos must be subjected to another MPI approved me thod of processing, listed in MPI-STD-
TVTL ; or 
c) The embryo donor must be subjected to a test for M. bovis listed in MPI-STD-TVTL , with negati ve 
results; or  
d) Each embryo collection for export to New Zealand must be tested using a validated test for M. bovis  
listed in MPI-STD-TVTL , with negative results.   
Ministry for Primary Industries   Page 29 5.19 Exotic Salmonella  spp. (salmonellosi s) 
5.19.1   Risk management options presented in the 2009 IRA  
(1) Require official certification that donors originate from farms on which outbreaks of salmonellosis have not 
occurred in the previous 3 years.  
(2) Require culture of semen or embryos or wash fluid sedimen t.  Where pathogenic Salmonella  spp. exotic to 
New Zealand is isolated, importation of germplasm could be prohibited.  
5.19.2   Discussion  
(3) Salmonella  spp. generally require higher levels of contamination than would be likely to be present in 
germplasm, and an oral route of entry, to be infective.    
(4) Historically , risk from contamination of imported bovine germplasm with  exotic  Salmonella  spp. has been 
accepted  due to the consequence being largely unknown.   
(5) It was concluded that measures specifically against salmonellosis should not be placed in the import 
health standard for bovine germplasm.  
5.19.3   Recommend ation  
(1) It is proposed that no measure s be placed in the standard specifically against exotic Salmonella  spp.  
 
 
 
  
Ministry for Primary Industries   Page 30 6 Appendi ces  
6.1 Appendix 1: Comparative Efficacy Study  
 
 
Ministry for Primary Industries   Page 31  
 
Ministry for Primary Industries   Page 32  
 
 
 
 
Ministry for Primary Industries   Page 33 6.2 Appendix 2: Antibiotic Treatment of Bovine Semen  
 
 
 
 
Ministry for Primary Industries   Page 34  
 
Ministry for Primary Industries   Page 35  
 
Ministry for Primary Industries   Page 36  
 
Ministry for Primary Industries   Page 37  
 
Ministry for Primary Industries   Page 38  
 
Ministry for Primary Industries   Page 39  
