Epidemiological investigation of norovirus infections in Punjab, Pakistan, through the One Health approach.

Introduction: Norovirus, mainly associated with acute gastroenteritis, is very contagious and can affect a vast range of species ranging from cattle, pigs, dogs, mice, cats, sheep, and lions to humans. It is a foodborne pathogen that mainly transmits through the fecal-oral route. Methods: This is the first-ever study conducted in Lahore and Sheikhupura districts of Punjab, Pakistan, to investigate noroviruses through the One Health approach. From January 2020 to September 2021, 200 fecal samples were collected from clinical cases of hospitalized patients and 200 fecal samples from sick animals at veterinary hospitals and local farms. In addition, 500 food and beverage samples were collected from street vendors and retail stores. A predesigned questionnaire was used to assess the risk factors and clinical characteristics of sick people and animals. Results and discussion: Overall, 14% of the human clinical samples were positive by RT-PCR for genogroup GII. All bovine samples were negative. Food and beverage samples were tested in pools, resulting in sugarcane juice samples positive for genogroup GII. Previous contact with acute gastroenteritis patients, sex, and presence of vomiting were found to be significant risk factors (p ≤ 0.05). The substantial number of diarrhea cases associated with noroviruses calls for additional studies to investigate the epidemiology and transmission and to improve surveillance.

Isolation and identification of Clostridium perfringens and its toxins from mutton in Lahore City, Punjab, Pakistan.

The present study was aimed to detect C. perfringens and identify its toxins in mutton samples collected from Lahore City in the Punjab Province of Pakistan. A total of 40 samples of minced and non­minced mutton were collected from local butcher and retail shops representing four areas of the city. The samples were subjected to ELISA for the detection of C. perfringens alpha, beta and epsilon toxins. The samples were simultaneously processed for bacterial isolation. The isolates were confirmed both by biochemical testing and a multiplex PCR targeting alpha, beta and epsilon toxin genes of C. perfringens. While 10% (4/40) of the samples were positive for C. perfringens alpha toxins, 17.5% (7/40) of the samples were positive for the alpha toxin gene. The present study indicated that the samples collected from the local butcher shops were contaminated with C. perfringens and its toxins. Interestingly, no such contamination was detected in any of the samples collected from retail meat shops. In conclusion, improper hygienic conditions at butcher shops could lead to the contamination of mutton with C. perfringens and its toxins.

Prevalence and molecular characterization of typhoidal and non-typhoidal Salmonella isolated from meat and environmental samples of retail shops of Lahore Punjab, Pakistan.

Non-typhoidal Salmonellae are important foodborne bacterial pathogens that can cause bacteremia, gastroenteritis, and subsequent infection. The aim of the study was to determine the prevalence of Salmonella in the live bird market and retail shops of Lahore (Pakistan). A total of 720 samples of chicken meat, chopping board, cages, hands, and transportation vans were collected. Salmonella was recovered from 103 (14.36%) samples. The highest prevalence (33.33%) was found in transportation van samples followed by chicken meat samples (17.26%). In the towns of Lahore, the highest prevalence was found in Samanabad Town (19%) followed by Data Ganj Bakhsh Town (17%) with the lowest in Gulberg Town (6.9%). Salmonella Typhimurium was most common (35.92%) followed by S. Enteritidis (25.24%), S. Dublin (14.56%), S. Gallinarum biovar Gallinarum (8.74%), and untyped Salmonella species (15.53%). This was the first baseline study of the prevalence of non-typhoidal Salmonella at the live bird market and retail shops of Lahore. Implementation of appropriate control measures is required at both the human side and poultry food production chain to reduce the burden and transmission of the zoonotic Salmonellae.

Effect of plasma progesterone on oocyte recovery, oocyte quality, and early in-vitro developmental competence of embryos in Bos indicus dairy cows.

The objective of present study was to determine the effect of plasma progesterone (P4) on oocyte recovery, oocyte quality, and early in-vitro developmental competence of embryos in Bos indicus dairy cows. The ovaries were collected in an abattoir. These ovaries (n = 750) were divided into two groups: 1) estrous CYCLIC (n = 318), and 2) estrous ACYCLIC (n = 432). Mean serum concentrations of P4 in a subset of (n = 85; 4.21 ± 0.4 ng/ml compared with 0.5 ± 0.2 ng/ml; P < 0.05) were greater in estrous CYCLIC as compared to ACYCLIC cows, respectively. The mean number of oocytes recovered per ovary (6.5 ± 0.5 compared with 4.0 ± 0.2; P < 0.05) was greater for estrous CYCLIC than ACYCLIC cows, respectively. The oocytes with grade I_+_II quality (55.3% compared with 47.6%; P < 0.05) were greater, whereas, there was lesser percentage with grade III_+_IV quality (44.5% compared with 52.4%; P < 0.05) from estrous CYCLIC as compared with ACYCLIC cows, respectively. Cleavage rate (70.9% compared with 52.8%; P < 0.05) was greater for embryos derived from estrous CYCLIC than ACYCLIC cows, respectively. Similarly, the embryo developmental rates to the 8- (38.5% compared with 20.8%; P < 0.05) and 16- (20.0% compared with 10.9%; P < 0.05) cell stage were greater for embryos derived from estrous CYCLIC as compared to ACYCLIC cows, respectively. In conclusion, the presence of greater plasma P4 has a beneficial effect on oocyte recovery, oocyte quality, and early IVEP outcomes in Bos indicus dairy cows.

Prevalence and Molecular Diagnosis of Babesia ovis and Theileria ovis in Lohi Sheep at Livestock Experiment Station (LES), Bahadurnagar, Okara, Pakistan.

BACKGROUND: Babesia ovis and Theileria ovis are among the important and main etiological agents causing ovine babesiosis and ovine theileriosis, causing severe economic losses among sheep and goats. The aim of the present study was to determine the prevalence and molecular diagnosis of B. ovis and T. ovis in Lohi sheep at Livestock Experiment Station Bahadurnagar, Okara, Pakistan. METHODS: The prevalence of B. ovis and T. ovis was investigated in 200 Lohi sheep of mixed age and sex by PCR during 2011. The assay was employed using primers Bbo-F & Bbo-R, specific for a 549-bp fragment in B. ovis genomic DNA and primers TSsr 170F & TSsr 670R, specific for a 520-bp fragment in T. ovis genomic DNA. The animals were also screened for both haemoparasites through stained thin blood smears. RESULTS: Thirty two (16%), 48 (24%) and 26 (13%) were the number of animals found positive for B. ovis, T. ovis and for mixed infection with both parasites, respectively, through microscopy. Sixty eight (34%), 73 (37%) and 42 (21%) were the number of animals found positive for B. ovis, T. ovis and for mixed infection with both parasites, respectively, through PCR test. CONCLUSION: The results indicate the high sensitivity of PCR for surveying babesiosis and theileriosis and there is noteworthy prevalence of these diseases in sheep at an experimental station where environmental conditions are relatively controlled as compared to field conditions.