Analyses of Extended-Spectrum-ß-Lactamase, Metallo-ß-Lactamase, and AmpC-ß-Lactamase Producing Enterobacteriaceae from the Dairy Value Chain in India.

The consumption of milk contaminated with antibiotic-resistant bacteria poses a significant health threat to humans. This study aimed to investigate the prevalence of Enterobacteriaceae producing ß-lactamases (ESBL, MBL, and AmpC) in cow and buffalo milk samples from two Indian states, Haryana and Assam. A total of 401 milk samples were collected from dairy farmers and vendors in the specified districts. Microbiological assays, antibiotic susceptibility testing, and PCR-based genotyping were employed to analyze 421 Gram-negative bacterial isolates. The overall prevalence of ß-lactamase genes was 10% (confidence interval (CI) (7-13)), with higher rates in Haryana (13%, CI (9-19)) compared to Assam (7%, CI (4-11)). The identified ß-lactamase genes in isolates were blaCMY, blaMOX, blaFOX, blaEBC, and blaDHA, associated with AmpC production. Additionally, blaCTX-M1, blaSHV, and blaTEM were detected as ESBL producers, while blaVIM, blaIMP, blaSPM, blaSIM, and blaGIM were identified as MBL producers. Notably, Shigella spp. were the dominant ß-lactamase producers among identified Enterobacteriaceae. This study highlights the presence of various prevalent ß-lactamase genes in milk isolates, indicating the potential risk of antimicrobial-resistant bacteria in dairy products. The presence of ß-lactam resistance raises concern as this could restrict antibiotic options for treatment. The discordance between genotypic and phenotypic methods emphasizes the necessity for comprehensive approaches that integrate both techniques to accurately assess antibiotic resistance. Urgent collaborative action incorporating rational and regulated use of antibiotics across the dairy value chain is required to address the global challenge of ß-lactam resistance.

Farmer's knowledge and suggested approaches for controlling aflatoxin contamination of raw milk in Pakistan.

Monitoring of aflatoxin levels in milk is often complicated in developing countries due to the dominance of informal markets channeling milk in raw form. Farmer's awareness and voluntary participation in aflatoxin mitigation can be critical in such scenarios. Therefore, the present study was conducted to understand the perceptions of dairy farmers about aflatoxins and link it with aflatoxin mitigation programs on milk in Pakistan. Information was collected from 450 peri-urban dairy farmers in seven cities using questionnaires. Majority (77.9%) of the farmers were aware of the negative impact of moldy feed on animal health. However, only 40.6% of the farmers were aware of the transferability of the toxins from moldy feed to milk. The farmers had almost no awareness of aflatoxins as 95% never heard of the term. After receiving an onsite briefing on effects of the toxin on animal and human health, and its transferability to milk, 98.3% farmers showed willingness to buy aflatoxin-safe feedstuffs, while 88.5% showed willingness to control aflatoxin in milk. Around half of the farmers considered aflatoxin control programs as affordable. On average, farmers agreed to pay 10.1% higher price for aflatoxin certified oilseed cakes. Availability of feedstuffs certified of low aflatoxin content was suggested by 22% of the participants as the critical step in reducing aflatoxins in milk. Other important suggestions included; subsidy on quality feeds (18%), raising awareness (18%), and legislation and monitoring (16%). The present results suggest that the current practice of milk monitoring in the country can yield desirable results only if it is coupled with feed certification programs ensuing availability of aflatoxin-safe feeds. Further, awareness can positively impact participation of producers in aflatoxin control programs. In this regard, awareness about effects of aflatoxins on animal health was found to be a more powerful trigger of voluntary control compared with the awareness of the toxin's transferability to milk.

Rabies Vaccination in Dogs in Laos: Owner Knowledge and Serological Status of Dogs.

Rabies is an infectious disease which is virtually 100% fatal. Humans are most often infected through the bite of an infected dog, and most cases could be prevented by vaccinating dogs. However, vaccination coverage is insufficient in most countries where canine rabies occurs endemically. This study conducted interviews and sampling of dogs in Laos to understand more about the barriers for vaccination and to evaluate the antibody status of dogs using a commercial ELISA. The study found that only 62% out of 359 dog owners knew what rabies was, and only 24% knew the disease could be fatal. Higher education was associated with higher knowledge scores. Only 56 out of 437 (13%) dogs had been rabies vaccinated according to their owner, and out of these dogs, only 34 (61%) had antibodies, and only 48% had adequate levels (above 0.5 IU/mL). However, 24% of the dogs with no known history of vaccination had antibodies, indicating either exposure or vaccination in the past without the owner's awareness. In conclusion, this study indicates that there is a low level of knowledge about rabies, and that owner knowledge is not a good indicator of whether a dog is vaccinated or not.

The Distribution and Composition of Vector Abundance in Hanoi City, Vietnam: Association with Livestock Keeping and Flavivirus Detection.

BACKGROUND: Dengue virus and Japanese encephalitis virus are two common flaviviruses that are spread widely by Aedes and Culex mosquitoes. Livestock keeping is vital for cities; however, it can pose the risk of increasing the mosquito population. Our study explored how livestock keeping in and around a large city is associated with the presence of mosquitoes and the risk of them spreading flaviviruses. METHODS: An entomological study was conducted in 6 districts with 233 households with livestock, and 280 households without livestock, in Hanoi city. BG-Sentinel traps and CDC light traps were used to collect mosquitoes close to animal farms and human habitats. Adult mosquitoes were counted, identified to species level, and grouped into 385 pools, which were screened for flaviviruses using a pan-flavivirus qPCR protocol and sequencing. RESULTS: A total of 12,861 adult mosquitoes were collected at the 513 households, with 5 different genera collected, of which the Culex genus was the most abundant. Our study found that there was a positive association between livestock keeping and the size of the mosquito population-most predominantly between pig rearing and Culex species (p < 0.001). One pool of Cx. tritaeniorhynchus, collected in a peri-urban district, was found to be positive for Japanese encephalitis virus. CONCLUSIONS: The risk of flavivirus transmission in urban areas of Hanoi city due to the spread of Culex and Aedes mosquitoes could be facilitated by livestock keeping.

Understanding Antibiotic Usage on Small-Scale Dairy Farms in the Indian States of Assam and Haryana Using a Mixed-Methods Approach-Outcomes and Challenges.

The use and misuse of antibiotics in both humans and animals contributes to the global emergence of antimicrobial resistant (AMR) bacteria, a threat to public health and infection control. Currently, India is the world's leading milk producer but antibiotic usage within the dairy sector is poorly regulated. Little data exists reflecting how antibiotics are used on dairy farms, especially on small-scale dairy farms in India. To address this lack of data, a study was carried out on 491 small-scale dairy farms in two Indian states, Assam and Haryana, using a mixed method approach where farmers were interviewed, farms inspected for the presence of antibiotics and milk samples taken to determine antibiotic usage. Usage of antibiotics on farms appeared low only 10% (95% CI 8-13%) of farmers surveyed confirmed using antibiotics in their dairy herds during the last 12 months. Of the farms surveyed, only 8% (6-11%) had milk samples positive for antibiotic residues, namely from the novobiocin, macrolides, and sulphonamide classes of antibiotics. Of the farmers surveyed, only 2% (0.8-3%) had heard of the term "withdrawal period" and 53% (40-65%) failed to describe the term "antibiotic". While this study clearly highlights a lack of understanding of antibiotics among small-scale dairy farmers, a potential factor in the emergence of AMR bacteria, it also shows that antibiotic usage on these farms is low and that the possible role these farmers play in AMR emergence may be overestimated.

Low Prevalence of Cysticercosis and Trichinella Infection in Pigs in Rural Cambodia.

Cysticercosis and Trichinella spp. infection are parasitic zoonoses prevalent among pigs in Southeast Asia, where pork is the most important source of meat. In rural Cambodia, many pigs are raised extensively in family backyards, and information regarding the prevalence in rural small-scale pig production is very limited. This study was conducted in four provinces in north-eastern Cambodia to determine the seroprevalence of porcine cysticercosis and Trichinella spp. infection in rural villages, and to identify possible risk factors. Only households with less than 10 pigs above three months old were eligible. In total, 139 households participated, and 242 blood samples were collected. Farmers were interviewed about food and hygiene habits, disease knowledge and practices. The serum samples were analysed by ELISA to determine antigens to Taenia spp. cysticerci or antibodies to Trichinella spp. muscle larvae. Positivity among the pigs was 11.2% (95% CI 7.5-15.8) for Taenia spp. cysts and 2.5% (95% CI 0.9-5.4) for Trichinella spp. Cysticerci were more common in the province Preah Vihear (p < 0.001) than in the other provinces. Risk factors associated with porcine cysticercosis were management systems for the pigs and access to human faeces (p < 0.001). Trichinella spp. infection in pigs was more common in the province Ratanakiri (p = 0.001). The main risk factor associated with Trichinella spp. transmission was feeding pigs with food waste (p = 0.048). More men had heard about cysticercosis than women (p = 0.002), and men also consumed undercooked pork meat to a greater extent (p = 0.004). Although the present study is relatively small, several risk factors could be identified for porcine infection with Taenia spp. and Trichinella spp., which can be used to guide future interventions to improve both porcine and human health in these provinces.

Leptospirosis in Ruminants in Yogyakarta, Indonesia: A Serological Survey with Mixed Methods to Identify Risk Factors.

Leptospirosis is a zoonotic disease occurring worldwide with reproductive symptoms and production losses in livestock, while humans can suffer fatal renal failure. In Yogyakarta Special Province, Indonesia, there have been several outbreaks with high case fatality, demonstrating the public health importance, but there is limited understanding of the epidemiology. This study used an EcoHealth approach to ensure transdisciplinarity and community participation. Seroprevalence of Leptospira in animals was studied between October 2011 and May 2013 in 15 villages. Serum samples from 1404 cattle and 60 small ruminants were screened by a Microscopic Agglutination Test (MAT), first in pools, and then the individual positive samples were identified. Focus group discussions including farmers, village officials, and official stakeholders were used to explore knowledge and behavior of zoonotic diseases, particularly leptospirosis. Two small ruminants were seropositive for Leptospira icterohemorrhagiae. From the cattle, 3.7% were seropositive, and the most common serovars were Leptospira hardjo, followed by L. icterohemorrhagiae. Out of all farms, 5.6% had at least one positive cattle. Risk factor analyses showed that the risk of the farm being seropositive increased if the farmer used water from an open source, or if farming was not the main occupation. This study showed the presence of Leptospira spp. in ruminants in Yogyakarta and identified use of open water as a risk factor for the livestock. We also observed that the knowledge related to leptospirosis was low, and risky farm management practices were commonly employed.

Aflatoxin Contamination of Milk Produced in Peri-urban Farms of Pakistan: Prevalence and Contributory Factors.

Aflatoxin M1 contamination of milk in Pakistan, like many developing countries, is poorly understood. The present study was therefore conducted to determine AFM1 contamination of milk and its contributory factors in Pakistan. We sampled milk and feedstuffs from 450 peri-urban dairy farms in seven major cities following a cross-sectional study design. Analysis of milk using ELISA revealed high contamination with an overall average of 3164.5 ng of AFM1/L, and significant differences (p < 0.001) between cities. The milk sampled from Gilgit, in northern hilly areas, had an average AFM1 level of 92.5 ng/L. Milk from other cities had 3529.7 ng/L average contamination, with only 5.7% samples qualifying the maximum tolerable limit of 500 ng of AFM1/L. Heavy mean aflatoxin contamination was found in bakery waste (724.6 µg/kg), and cottonseed cake (600.8 µg/kg). Rest of the other feedstuffs had moderate to low mean aflatoxin contamination, ranging from 66.0 µg/kg in maize stover to 3.4 µg/kg in wheat bran. The mean aflatoxin level in commercial dairy concentrates was 32.7 µg/kg. About 80% of the total aflatoxin intake of dairy animals was contributed by cottonseed cake alone due to its high aflatoxin contamination and proportion in dairy rations. On-farm storage time of oilseed cakes varied (p < 0.01) in different cities but was not associated with aflatoxin contamination. The exceptionally high AFM1 contamination suggests that milk from peri-urban dairy farms is a serious public health threat in Pakistan. This situation can be mitigated by reducing aflatoxin contamination in cottonseed cake and promoting the use of commercial concentrates and other feedstuffs with low contamination.

Aflatoxin M1 levels in different marketed milk products in Nairobi, Kenya.

Milk is an important source of energy and nutrients, especially for children, and in Kenya, milk consumption is higher than other countries in the region. One major concern with milk is the risks of chemical contaminants, and reports of high levels of aflatoxin M1 (AFM1) in milk in Kenya has been causing public health concerns. This study collected marketed milk products every month during 1 year, just as a consumer would purchase them from retailers and traders in a low-income area, and a major supermarket in a middle/high-income area. In total, 291 sampled milk products (raw, pasteurised, UHT milk, yoghurt and lala) were collected and analysed for AFM1 using a commercial ELISA kit. More than 50% of the samples exceeded 50 ng/kg (the level allowed in the EU), but only three samples exceeded 500 ng/kg (the level allowed in the USA). Geometric mean AFM1 level was 61.9 ng/kg in the 135 samples from the low-income area while it was 36.1 ng/kg in the 156 from the higher income area (p < 0.001). The levels varied significantly depending on the time of year, with lowest levels of milk in January. There were also differences between manufacturers and products, with UHT milk having lower levels. There was no difference depending on the price for all dairy products, but when only including milk, higher price was associated with lower levels of AFM1. In conclusion, this study shows that milk purchased by a consumer is likely to contain AFM1 above 50 ng/kg, and that further research is needed to find ways to mitigate AFM1 contamination through working with farmers and milk processors both in the formal and informal sectors.