Bacterial Metabarcoding of Raw Palm Sap Samples from Bangladesh with Nanopore Sequencing.

The traditional practice of harvesting and processing raw date palm sap is not only culturally significant but also provides an essential nutritional source in South Asia. However, the potential for bacterial or viral contamination from animals and environmental sources during its collection remains a serious and insufficiently studied risk. Implementing improved food safety measures and collection techniques could mitigate the risk of these infections. Additionally, the adoption of advanced food analytical methods offers the potential to identify pathogens and uncover the natural bacterial diversity of these products. The advancement of next-generation sequencing (NGS) technologies, particularly nanopore sequencing, offers a rapid and highly mobile solution. In this study, we employed nanopore sequencing for the bacterial metabarcoding of a set of raw date palm sap samples collected without protective coverage against animals in Bangladesh in 2021. We identified several bacterial species with importance in the natural fermentation of the product and demonstrated the feasibility of this NGS method in the surveillance of raw palm sap products. We revealed two fermentation directions dominated by either Leuconostoc species or Lactococcus species in these products at the first 6 h from harvest, along with opportunistic human pathogens in the background, represented with lower abundance. Plant pathogens, bacteria with the potential for opportunistic human infection and the sequences of the Exiguobacterium genus are also described, and their potential role is discussed. In this study, we demonstrate the potential of mobile laboratory solutions for food safety purposes in low-resource areas.

Snapshot study of canine distemper virus in Bangladesh with on-site PCR detection and nanopore sequencing.

Canine distemper virus (CDV) is a highly contagious virus that affects domestic and wild animals, causing severe illness with high mortality rates. Rapid monitoring and sequencing can provide valuable information about circulating CDV strains, which may foster effective vaccination strategies and the successful integration of these into conservation programs. During two site visits in Bangladesh in 2023, we tested a mobile, deployable genomic surveillance setup to explore the genetic diversity and phylogenetic patterns of locally circulating CDV strains. We collected and analysed 355 oral swab samples from stray dogs in Rajshahi and Chattogram cities, Bangladesh. CDV-specific real-time RT-PCR was performed to screen the samples. Out of the 355 samples, 7.4% (10/135) from Rajshahi city and 0.9% (2/220) from Chattogram city tested positive for CDV. We applied a real-time RT-PCR assay and a pan-genotype CDV-specific amplicon-based Nanopore sequencing technology to obtain the near-completes. Five near-complete genome sequences were generated, with phylogenetic relation to the India-1/Asia-5 lineage previously identified in India. This is the first study to provide genomic data on CDV in Bangladesh and the first demonstration of a mobile laboratory setup as a powerful tool in rapid genomic surveillance and risk assessment for CDV in low resource regions.

Molecular identification, antimicrobial resistance and virulence gene profiling of Staphylococcus spp. associated with bovine sub-clinical mastitis in Bangladesh.

This study was conducted to investigate the diversity and antimicrobial resistance profiling of Staphylococcus species causing sub-clinical mastitis (SCM) in dairy herds in Bangladesh as well as putative risk factors associated with the infections. Individual quarter milk samples were collected from a total of 284 lactating cows from 30 dairy farms were screened by means of California mastitis test; 178 (62.7%) of them had at least of quarter affected by SCM. After conventional microbiological isolation procedures, PCR tests were used for Staphylococcus species identification and detection of antimicrobial resistance and virulence genes. S. chromogenes (65.7%) was the most predominant species followed by, S. epidermidis (20.2%), S. haemolyticus (19.1%), S. aureus (15.7%), and S. sciuri (5.6%). High levels of antimicrobial resistance to ampicillin and amoxicillin/clavulanic acid were observed in S. aureus (82.1% and 75%) and S. sciuri (80% and 70%), while resistance to cefepime was markedly higher in S. chromogenes (95.7%), S. haemolyticus (94.1%), and S. epidermidis (97.2%). Multidrug resistance isolates were identified in all five species. The mecA gene was detected in S. aureus (32.1%) and S. chromogenes (5.98%). In addition, 20% S. sciuri and 17.7% S. haemolyticus carried the cytotoxin (pvl) gene, while 14.3% S. aureus harbored the toxic shock syndrome toxin (tst) gene. Multivariable logistic regression analysis identified "Old aged" (OR [CI]: 3.5 [1-12.4]); "Early stage of lactation" (OR [CI]: 3.4 [1.2-9.7]) and, "Firm udder condition" (OR [CI]: 4.2 [1.2-14.6]) as risk factors associated with SCM caused by S. aureus, S. chromogenes, and S. haemolyticus, respectively. Moreover, "Use of antimicrobials" (OR [CI]: 10.4 [3.4-32.1] and "History of previous clinical mastitis" (OR [CI]: 4.9 [1.2-19.7] for the carriage of methicillin-resistant Staphylococcus spp.

Clinico-epidemiological investigation of feline panleukopenia and parvoviral enteritis in the two largest pet hospitals in Bangladesh.

OBJECTIVE: A clinico-epidemiological study was conducted at two veterinary hospitals in Dhaka to evaluate the prevalence of parvoviral enteritis (PVE) in dogs and feline panleukopenia (FPL) in cats, to detect factors associated with them, and to identify their common clinical signs. MATERIALS AND METHODS: Clinico-epidemiological data were collected for a total of 88 dogs and 129 cats which were presented to the hospitals, entered into MS excel 2010, and then transferred to STATA-14 software for conducting descriptive and univariable statistical analyses (Fisher's exact test). The maps showing the spatial distribution of PVE and FPL were produced by using QGIS software version 2.18.13. RESULTS: PVE in dogs and FPL in cats had the highest prevalence compared to other diseases (34.1%; 95% confidence interval: 24.3-44.9; N = 88 and 20.2%; 95% confidence interval: 13.6-28.1; N = 129, respectively). Young age (43.8%), poor body condition (58.4%), and exotic breeds (44.2%) were significantly associated with the occurrence of PVE (p ≤ 0.05). Poor body condition (27.8%) and non-vaccination status (26.7%) were associated with FPL (p ≤ 0.05). Both PVE and FPL showed clinical signs of being off-feed, frequent vomiting, weakness, and moderate dehydration. CONCLUSION: The high prevalence of PVE and FPL indicates that they are common in dogs and cats in Dhaka, Bangladesh. The factors related to PVE are young age, exotic breed, and poor body condition score (BCS), and FPL are poor BCS and non-vaccination. Specific measures such as care during younger age, good nutrition, and routine vaccination are needed to prevent and control PVE and FPL in Dhaka's dog and cat population.

Hematological and biochemical reference values of Asian house shrews (Suncus murinus) in Bangladesh.

BACKGROUND AND AIM: Determining reference values for hematological and biochemical parameters of Asian house shrew (Suncus murinus) is important for wildlife research to protect human health in surrounding communities. This study aimed to establish the reference values for selected hematology and serum clinical chemistry analyses that may contribute to research on shrew in future. MATERIALS AND METHODS: Blood samples (n=51) were collected from shrews between July and December 2015, Bangladesh, to estimate the levels of hemoglobin (Hb), packed cell volume (PCV), total leukocyte count (TLC), total erythrocyte count (TEC), lymphocyte, monocyte, neutrophil, eosinophil, basophil, calcium, phosphorus (P), sodium (Na), chloride (Cl), urea, glucose, total protein (TP), creatinine, and alanine transaminase (ALT). RESULTS: Although the values did not differ significantly among sexes, age was found to be a significant factor. Hb, PCV, TEC, glucose, and P were higher in males; eosinophil, Na, Cl, TP, and ALT were higher among females. Adults had significantly greater urea and glucose (p<0.05) while juveniles had insignificantly higher values for TLC, PCV, neutrophil, P, and TP. CONCLUSION: This study provides the first reference values for this species in Bangladesh and can be used to guide wildlife research studies.

Diversity and prevalence of parasitic infestation with zoonotic potential in dromedary camel ( Camelus dromedarius ) and fat-tailed sheep (dhumba) in Bangladesh.

OBJECTIVE: Parasitic infestation is a major cause of losses in livestock production in tropical regions. A cross-sectional study was conducted to determine the prevalence of Gastro-intestinal (GI) parasites of dromedary camel (Camelus dromedarius) and fat-tailed sheep (dhumba), and the prevalence of hemoparasites in camel from Dhaka, Bangladesh. MATERIALS AND METHODS: A total of 87 fecal samples (32 dhumba and 55 camel) and 55 camel blood samples were collected during September-October 2015. Fecal samples were examined by direct smear, sedimentation method, flotation technique, and McMaster technique for GI parasite. Giemsa stained blood smears were examined under microscope for hemoparasite detection. RESULTS: 62% camel (n = 34; 95% confidence interval (CI): 47.7-74.6) were infected with at least one genus of parasite. 15% camel were harboring more than one genus of parasite. The prevalence of GI parasite and hemoparasite in camel were recorded as Trichuris spp. (n = 16; 29%; 95% CI: 17.6-42.9), Balantidium coli (n = 12; 22%; 95% CI: 11.8-35.0), Trichostrongylus spp. (n = 7; 13%; 95% CI: 5.3-24.5), Strongyloides spp. (n = 5; 9%; 95% CI: 3.0-20.0), Anaplasma spp. (n = 5; 9%; 95% CI: 3.02-20.0), Paragonimus spp. (n = 1; 2%; 95% CI: 0.05-9.7), Schistosoma spp. (n = 1; 2%; 95% CI: 0.05-9.7), Hymenolepis spp. (n = 1; 2%; 95% CI: 0.05-9.7), Moniezia spp. (n = 1; 2%; 95% CI: 0.05-9.7), and Babesia spp. (n = 1; 2%; 95% CI: 0.05-9.7). Mean EPG feces of camel was 291.76 ± 42.03 with a range of 0-1,400. Total 59.4% dhumba (n = 19; 95% CI: 41-76) were positive for GI parasite, including Trichostrongylus spp. (n = 10; 31.3%; 95% CI: 16.1-50), Strongyloides spp. (n = 9; 28%; 95% CI: 13.8-46.8), B. coli (n = 5; 15.6%; 95% CI: 5.3-32.8), and Trichuris spp. (n = 4; 12.5%; 95% CI: 3.5-28.9). CONCLUSIONS: High percentage of parasitic infestation in camel and dhumba in the present study refers to the necessity of use of anthelmintic for health and production improvement and to prevent zoonotic parasite transmission to animal handler and workers.

Risk factors and therapy for goat mastitis in a hospital-based case-control study in Bangladesh.

Bangladesh has a large population of goats, which contribute to the income, nutrition and welfare of the households of many families. Mastitis in goats has a low incidence, but is often very severe, making veterinary care necessary. The aim of this study was to identify seasonality and risk factors for goat mastitis in a hospital-based matched case-control study in a teaching veterinary hospital in Chittagong, Bangladesh and to describe the range of antimicrobial treatments applied in this situation. Cases of mastitis and controls were drawn from the hospital patient recording system, along with their risk factor status. Multiple imputation was applied to deal with the missing values in the data analysis. Mastitis occurred somewhat more in the rainy season, and comprised about 3% of all goats admitted to the hospital during January 2011-June 2014. Free-ranging farming system, poor body condition score and non-native goat breeds were significantly associated with case status. Treatment of clinical mastitis was variable and unsystematic, but the use of gentamicin was commonly recorded. The need for more prudent and evidence-based antimicrobial therapies is discussed.