Trypanosomiasis is one of the severe pathogenic infections, caused by several Trypanosoma species, affecting both animals and humans, causing substantial economic losses and severe illness. The objective of this study was to determine the molecular diagnosis and the risk factors associated with trypanosomiasis in District Jhang, Punjab, Pakistan. For this purpose, blood samples were randomly collected from 200 horses. A predesigned questionnaire was used to collect data on risk factors before the sample collection. The microscopy examination through Giemsa staining, formol gel test and PCR techniques were used to find the prevalence. The prevalence was recorded as 22.5% with microscopy examination, 21% through formol gel test and 15.5% with PCR based results. Analysis of risk factors associated with Trypanosoma brucei evansi occurrence was carried out using Chi-square test. It showed the prevalence of Trypanosoma brucei evansi was significantly (p<0.05) associated with sex, age, rearing purpose and body condition whereas non-significantly (p>0.05) with insects control practices. This study supports the idea that PCR is a sensitive, robust and more reliable technique to diagnose trypanosomiasis. It was concluded that Trypanosoma brucei evansi is widely prevalent in Jhang (Pakistan), highlighting a dire need to develop control strategies and education programmes to control this disease in developing countries.

Members of Enterobacteriaceae are known to produce extended-spectrum beta-lactamases (ESBL) which hydrolyze the beta-lactam group of antibiotics. The existence of ESBL-producing Salmonella enterica ( S. enterica) and Escherichia coli ( E. coli) harbored by urban avifauna was investigated in this study. Dropping samples (n= 180) were collected from six different bird species in the district Jhang, Punjab province, Pakistan. Isolation and identification of ESBL isolates were made by using cefotaxime- (4 mg/L) supplemented MacConkey agar and double disc synergy test (DDST). Polymerase chain reaction (PCR) was performed for the detection of four different ESBL genes including bla _CTX-M, bla _TEM, bla _SHV and bla _OXA. A total of 42.69% isolates were confirmed as ESBL via DDST including 30.64% S. enterica and 49.54% E. coli. The incidence of ESBL S. enterica and ESBL E. coli was found highest in egret ( Ardea alba) and pigeon ( Columba livia) as 64.28% and 78.95%, respectively. The bla _CTX-M gene was detected in 57.89% and 64.81% of isolates of S. enterica and E. coli, respectively. Among other genes in S. enterica and E. coli, bla _TEM (21.05%, 20.4%); bla _SHV (15.78%, 9.26%), and bla _OXA (5.26%, 5.56%) were detected, respectively. All of the tested isolates were found resistant to at least one of the thirteen antimicrobial agents except meropenem. To the best of our knowledge, this is the first study reporting the incidence and genetic diversity of ESBL bacteria associated with urban avifauna in Pakistan. The urban avifauna can serve as a potential subject of bio-surveillance to monitor the emergence of antimicrobial-resistant bacteria.