Simple SummaryMexico is an important honey producer, but not much information exists about the health of honey bees in the country. This study analyzed the sanitary status of adult honey bees in five different beekeeping regions of Mexico. Samples from hives were analyzed to identify pests, parasites, and viruses. The miteVarroa destructorwas found in 83.5% of the samples, with the Pacific Coast having the highest frequency (>95%) and levels (4.5% ± 0.6). Another mite,Acarapis woodi, was found in only one sample from the Pacific Coast, whereas the fungiNosemaspp. were present in 48.5% of the samples, with the highest frequency in the Yucatan Peninsula (64.6%). For viruses, deformed wing virus (DWV) was more frequently found in the Pacific Coast region (44.7%), and Israeli acute paralysis virus (IAPV) was detected in only 3.2% of the samples. Sacbrood bee virus (SBV) was frequently found in the High Plateau region (36.4%), and the small hive beetle (SHB) was common in the Yucatan Peninsula (39.2%). This information could be useful to design disease control strategies for honey bee colonies in different regions of Mexico.AbstractMexico is a major honey producer, but not much information exists about the health status of honey bees (Apis melliferaL.) in the country. This study was conducted to determine the sanitary status of adult honey bees in Mexico’s five beekeeping regions. Samples from 369 apiaries were diagnosed to identify pathogens such asVarroa destructor, which was quantified,Acarapis woodi,Nosemaspp., and five viruses. Colonies were also inspected for the presence of the small hive beetle (SHB),Aethina tumida.Varroa destructorwas found in 83.5% of the apiaries, with the Pacific Coast region having the highest prevalence (>95%) and rates (4.5% ± 0.6).Acarapis woodiwas detected in only one apiary from the Pacific Coast, whereasNosemaspp. were prevalent in 48.5% of the apiaries, with the highest and lowest frequencies in the Yucatan Peninsula and North regions (64.6% and 10.2%, respectively). For viruses, deformed wing virus (DWV) was detected in 26.1% of the apiaries, with the highest frequency in the Pacific Coast region (44.7%). Israeli acute paralysis virus (IAPV) was diagnosed in 3.2% of the samples and sacbrood bee virus (SBV) in 23.3% of them, with the highest frequency in the High Plateau region (36.4%). Chronic bee paralysis and Kashmir bee viruses were not detected. SHB prevalence was 25.2% nationwide, with the highest frequency in the Yucatan Peninsula (39.2%). This study shows that the most common parasites of adult honey bees in Mexico areV. destructorandNosemaspp., and that the most prevalent virus is DWV, whereas SHB is highly prevalent in the Yucatan Peninsula. This information could be useful to design disease control strategies for honey bee colonies in different regions of Mexico.