Sentinel surveillance for influenza A viruses in Lahore District Pakistan in flu season 2015-2016.

BACKGROUND: Influenza A virus (IAV) remains an important global public health threat with limited epidemiological information available from low-and-middle-income countries. The major objective of this study was to describe the proportions, temporal and spatial distribution, and demographic and clinical characteristics of IAV positive patients with influenza like illness (ILI) and severe acute respiratory illness (SARI) in Lahore, Pakistan. METHODS: Prospective surveillance was established in a sentinel hospital from October 2015 to May 2016. All eligible outpatients and inpatients with ILI or SARI were enrolled in the study. Nasal and/or throat swabs were collected along with clinico-epidemiological data. Samples were tested by real-time RT-PCR (rRT-PCR) to identify IAV and subtype. The descriptive analysis of data was done in R software. RESULTS: Out of 311 enrolled patients, 284 (91.3%) were ILI and 27 (8.7%) were SARI cases. A distinct peak of ILI and SARI activity was observed in February. Fifty individuals (16%) were positive for IAV with peak positivity observed in December. Of 50 IAV, 15 were seasonal H3N2, 14 were H1N1pdm09 and 21 were unable to be typed. The majority of IAV positive cases (98%) presented with current or history of fever, 88% reported cough and 82% reported sore throat. The most common comorbidities in IAV positive cases were hepatitis C (4%), obesity (4%) and tuberculosis (6%). The highest incidence of patients reporting to the hospital was seen three days post symptoms onset (66/311) with 14 of these (14/66) positive for IAV. CONCLUSION: Distinct trends of ILI, SARI and IAV positive cases were observed which can be used to inform public health interventions (vaccinations, hand and respiratory hygiene) at appropriate times among high-risk groups. We suggest sampling from both ILI and SARI patients in routine surveillance as recommended by WHO.

Risk Factors of Influenza-Associated Respiratory Illnesses Reported to a Sentinel Hospital of Lahore, Pakistan: 2015-2016.

Epidemiological data about determinants of influenza A virus (IAV) in the Pakistani population is scarce. We aimed to conduct a prospective hospital-based active surveillance study from October 2015 to May 2016 to identify potential risk factors associated with IAV infection among patients with influenza-like illness (ILI) and severe acute respiratory illness (SARI). Surveillance was conducted in Lahore General Hospital, selected as a sentinel site in Lahore District, Pakistan. Nasal/throat samples were collected along with epidemiological and clinical data from enrolled patients. Real-time reverse-transcription polymerase chain reaction (rRT-PCR) was performed to identify IAV and its subtypes (H1N1pdm09, H3N2). Data were analyzed to determine risk factors and risk markers associated with IAV infections. A total of 311 suspected ILI and SARI cases were enrolled in the study, and among these 50 were IAV-positive. Of these 50 confirmed cases of IAV, 14 were subtyped as H1N1pdm09 and 15 were H3N2; the remaining 21 were untyped. A final multivariable model identified four independent risk factors/markers for IAV infection: exposure history to ILI patients within last 7 days and gender being male were identified as risk factors of IAV infection, while use of antibiotics prior to hospital consultation and presence of fever were identified as risk markers. We concluded that adopting nonpharmaceutical interventions like hand hygiene, masks, social distancing, and where possible, avoiding identified risk factors could decrease the risk of IAV infection and may prevent imminent outbreaks of IAV in the community.

Diagnostic accuracy of panoramic radiography and MRI for detecting signs of TMJ degenerative joint disease.

The objective of this study was to determine the diagnostic accuracy of panoramic radiography and magnetic resonance imaging (MRI) for detection of signs of temporomandibular joint (TMJ) degenerative joint disease (DJD). Panoramic radiography and bilateral MRI and computed tomography (CT) of the TMJs were performed for 705 subjects. Three calibrated board-certified radiologists who were blinded to the clinical findings interpreted all images. The diagnoses of DJD established using the panoramic radiographs and MRIs were compared to the reference standard diagnoses derived from the CTs. DJD was defined as the presence of at least 1 of the following 4 signs: a subcortical cyst, surface erosion, osteophyte formation, or generalized sclerosis. The target values for sensitivity and specificity were 70% or greater and 95% or greater, respectively. Compared to the reference standard CTs, the panoramic radiographs had the following sensitivity and specificity values: subcortical cysts, 14% and 100%, respectively; erosion, 20% and 100%, respectively; osteophytes, 12% and 100%, respectively; and sclerosis, 33% and 100%, respectively. The MRIs achieved the following sensitivity and specificity values: subcortical cysts, 32% and 100% respectively; erosion, 35% and 99% respectively; osteophytes, 71% and 98%, respectively; and sclerosis, 50% and 100%, respectively. The radiologists' interexaminer reliability was slight (κ = 0.16) when using panoramic radiographs, moderate (κ = 0.47) when using MRIs, and substantial when using CTs (κ = 0.71) for diagnosis of signs of DJD. Panoramic radiographs and MRIs had below-target sensitivity but above-target specificity in detecting all CT-depicted signs of DJD with the exception of osteophytes, for which MRIs demonstrated adequate diagnostic accuracy. Use of CT for diagnosis of TMJ DJD is recommended to avoid the false-negative findings that can occur if panoramic radiographs and MRIs are used.

Risk factors associated with Mycobacterium bovis skin positivity in cattle and buffalo in Peshawar, Pakistan.

A cross-sectional study was carried out to determine the prevalence of bovine tuberculosis (bTB) and associated risk factors in cattle and buffalo in Peshawar, Pakistan. Cattle and buffalo, randomly selected from all four towns of District Peshawar, were screened for bovine tuberculosis using comparative cervical intradermal tuberculin test (CCIT). For obtaining data on risk factors, sociodemographic condition, animal characteristics, and management, interviewer administered pretested questionnaire to animal owners. Multivariable logistic regression models were used to measure association between risk factors and comparative cervical intradermal tuberculin reactors. A total of 556 cattle and buffalo were screened for bovine tuberculosis. Out of 556 animals screened, 5.75 % (3.9-8.0 %) were found positive. The prevalence was higher in old animals (p = 0.001) as compared to younger animals. Prevalence also varied with source of animal (either raised on farm or purchased), stay of animals at night (indoor or outdoor), and herd size. Farmer's knowledge about transmission of TB from animals to human as well as signs and symptoms of TB was extremely low. Only 3.6 % farmers correctly stated the combination of three major symptoms of TB. Results of the study call for immediate intervention to control bTB in animals as well as its transmission to human population. Furthermore, it is suggested to emphasize on local epidemiology of bTB and husbandry practices of cattle and buffalo during the control program.

Toxinotyping and antimicrobial susceptibility of enterotoxigenic Clostridium perfringens isolates from mutton, beef and chicken meat.

A total of 300 meat samples comprising mutton, beef, and chicken meat (n = 100) collected from either local butcher shops or large meat outlets situated at various areas of Lahore City located in Punjab province of Pakistan were tested for the isolation of Clostridium perfringens. Prevalence of the organism was highest in the chicken (6 %) followed by mutton (5 %) and beef (1 %). Contamination level was high (10/150) in the samples collected from local butcher shops in comparison to the samples collected from large meat outlets (2/150). All of the raw meat samples were negative for the presence of alpha, beta and epsilon toxins of C. perfringens as detected through ELISA. Out of a total number of 12 isolates only half were capable of producing enterotoxins when cultured in trypticase glucose yeast (TGY) broth. Toxinotyping of the isolates showed that 3 were of type A while one each of the remaining three belonged to type B, C, and D. Antibiotic susceptibility testing of the toxin producing isolates revealed that C. perfringens were susceptible to chloramphenicol, ciprofloxacin, metronidazole, and ceftriaxone. All of the other drugs were relatively less effective with a least activity of amoxicillin against the isolates.

Investigation of equine influenza virus in two geographical regions of Pakistan.

PURPOSE: The present study was an attempt to elucidate the seroprevalence of equine influenza virus (H3N8) in two geographically distinct regions of Pakistan where vaccination is not routinely practiced. METHODS: A total number of 315 animals of family Equidae were included in the survey. Blood samples and nasal swabs were collected from the same animal and analyzed through ELISA and Hemagglutination Inhibition. RESULTS: The seroprevalence for EIV was 10 and 8.39 in districts under study. Out of 29 ELISA positive sera, 7 (24.1 %) showed antibodies against H1 and 22 (75.9 %) showed against H3 of influenza strains. Specie-wise antibody titer against H1 was 7.5 and 9.0 for horses and donkeys, respectively, while mules were negative. Similarly, antibody titer against H3 was 7.12, 6.9, and 6.0 for horses, donkeys, and mules, respectively. No strain was isolated from the nasal swabs despite very high antibody titers of H1 and H3 in the sera of the same animals. CONCLUSIONS: Influenza viruses are circulating in equine species and continuous surveillance is needed to keep check on any future outbreak.

Recommendations for Imaging of the Temporomandibular Joint. Position Statement from the American Academy of Oral and Maxillofacial Radiology and the American Academy of Orofacial Pain.

This position statement was developed by an ad hoc committee of the American Academy of Oral and Maxillofacial Radiology and the American Academy of Orofacial Pain. The committee reviewed the pertinent literature and drafted recommendations for imaging. This joint statement provides evidence-based recommendations and clinical guidance for applying appropriate diagnostic imaging to evaluate the temporomandibular joint (TMJ). This manuscript guides the design of TMJ imaging examinations, addresses in-office CBCT imaging, and provides timely evidence-based recommendations to evaluate the TMJ bony components, also addressing the use of MRI and other modalities to evaluate TMJ involvement in different pathologic conditions.

Assessment of aflatoxin M1 exposure and associated determinants in children from Lahore, Pakistan.

Aflatoxins are potent carcinogenic and immunomodulatory mycotoxins, and exposure may lead to deleterious effects on human health. This study aimed to detect aflatoxin M1 (AFM1) as biomarker of exposure and determine associated risk factors in children attending a specialized-childcare hospital in Lahore. Urine samples collected from 238 children (1-11 years) during winter (January-mid-March 2020) and hot-humid summer (August-September 2020) were tested for AFM1 presence using ELISA. Data on potential risk factors were also collected. Of 238 samples, 156 (65.5%) were positive for urinary AFM1. Season was significantly associated (OR = 2.64; 95% CI = 1.49-4.79; p = 0.001) with AFM1 positivity; prevalence was higher in hot-humid months (74.6%) than winter (57.3%). The place of living was also significantly associated (OR = 2.21; 95% CI = 1.25-3.97; p = 0.007), and urinary AFM1 positivity was higher in urban children (71.1%) compared to rural (58.3%). Median value for creatinine-adjusted AFM1 was 1.9 ng/mg creatinine (Q1-Q3 = 0.82-6.0 ng/mg creatinine), while non-creatinine-adjusted AFM1 was 0.57 ng/mL (Q1-Q3 = 0.23-1.4 ng/mL). Significantly higher urinary AFM1 levels were detected in children; age ≤2 years (p = 0.037), who consumed more milk (p = 0.048), and who presented to the nutrition clinic (p = 0.003). These findings highlight the need for an effective control program to reduce the AFM1 burden in children.

Intraoral salivary lymphoepithelial carcinoma: clinicopathologic and immunophenotypic characterization of 3 cases indicates elevated programmed death-ligand 1 expression.

OBJECTIVE: Intraoral salivary lymphoepithelial carcinoma (ISLEC) is a rare malignancy with programmed death-ligand 1 (PD-L1) expression levels that have been greatly understudied. We examined the clinicopathologic and immunophenotypic characteristics, including PD-L1 levels, of 3 cases of ISLEC. STUDY DESIGN: We searched the archives of 2 oral and maxillofacial pathology laboratories for specimens diagnosed as ISLEC between 1985 and 2022. We collected patient demographic and clinical data. Immunostaining for AE1/AE3, CK7, CD3, CD20, p16, p53, Ki67, and PD-L1 (SP263), as well as Epstein-Barr virus-encoded small RNAs (EBER) in situ hybridization (ISH) were performed. RESULTS: All 3 cases affected males aged 42 to 84 years (median = 61y) and involved the floor of the mouth, soft palate/uvula, and tongue. The lesions showed diffuse infiltration by non-keratinizing sheets and islands of undifferentiated carcinoma cells with associated dense lymphoplasmacytic inflammation. Immunohistochemically, all tumors showed AE1/AE3 positivity, selective p53 staining, and negativity for CK7 and p16. Ki67 highlighted 20%-80% of lesional cells. The inflammatory infiltrate comprised a mixed population of T and B lymphocytes. EBER ISH was positive in one case. All ISLECs displayed membranous, focal-to-diffuse, PD-L1 staining with tumor proportion score > 95% in two and 40-50% in the third case. CONCLUSIONS: The clinicopathologic and immunophenotypic characteristics of the cases we examined highlight the rarity of ISLEC and indicate overall high PD-L1 levels in this type of malignancy, rendering patients with ISLEC potential candidates for targeted α-PD-1/PD-L1 immunotherapy.

Prevalence of multi-drug resistant Pseudomonas aeruginosa isolated from selected residential sewages in Dutsin-Ma, Katsina State, Nigeria.

The global surge in Multidrug resistant (MDR) bacteria is an issue of great concern. Pseudomonas aeruginosa has been implicated in several nosocomial infections, where it has caused grave complications in immunocompromised patients. This is the first study to report the prevalence of MDR P. aeruginosa isolated from residential sewage in Dutsin-Ma, Katsina State, Nigeria. Pseudomonads count, isolation, biochemical characterization and antibiogram were carried out using standard microbiological procedures. This study examined sixty (60) samples from selected residential sewage in the study site collected at different intervals between July and September 2021. A total of 40 (66.7%) P. aeruginosa were isolated from the analyzed sewage samples. The highest (2.84x104) pseudomonad count was recorded from sewage samples collected from Kadangaru. Pseudomonas aeruginosa isolates from this sample site showed the highest (100%) resistance to cephalosporins (cefuroxime) and nitrofurantoin. Similarly, isolates from Miami area also demonstrated the highest (95%) resistance to a cephalosporin (ceftazidime). All (100%) isolates used in this study showed MDR resistance to tested antibiotics. The occurrence of MDR P. aeruginosa from a residential sewage site that may contaminate drinking water sources in the study area is of public health threat to the inhabitants. Surveillance and molecular epidemiology of antibiotics resistant bacteria are urgently needed in the study area.

Dosimetry of a cone-beam computed tomography machine compared with a digital x-ray machine in orthodontic imaging.

INTRODUCTION: Cone-beam computed tomography (CBCT) has become a routine imaging modality for many orthodontic clinics. However, questions remain about the amount of radiation patients are exposed to during the scans. This study determined the amounts of radiation potentially absorbed by a patient during orthodontic imaging with a CBCT machine with various scan settings compared with a conventional 2-dimensional digital x-ray machine. METHODS: The radiation exposures delivered by a next generation i-CAT CBCT machine (Imaging Sciences International, Hatfield, Pa) at various scan settings and orthopantomograph OP100/OC100 digital x-ray machine (Instrumentarium Dental, Tuusula, Finland) during panoramic and cephalometric radiography were recorded using thermoluminescent dosimeters placed inside a head and neck phantom. The manufacturer-recommended settings for an average adult male were used for both types of machines. Effective doses were calculated using the tissue-weighting factors recommended by the 2007 International Commission on Radiological Protection. RESULTS: The effective doses at various voxel sizes and field of view settings ranged from 64.7 to 69.2 µSv for standard resolution CBCT scans (scan time 8.9 s) and 127.3 to 131.3 µSv for high resolution full field of view scans (scan time 17.8 s), and measured 134.2 µSv for a high-resolution landscape scan with a voxel size as would be used for SureSmile (OraMetrix, Richardson, Tex) therapy (scan time 26.9 s). The effective doses for digital panoramic and lateral cephalometric radiographs measured 21.5 and 4.5 µSv, respectively. CONCLUSIONS: CBCT, although providing additional diagnostic and therapeutic benefits, also exposes patients to higher levels of radiation than conventional digital radiography.

Recommendations for imaging of the temporomandibular joint. Position statement from the American Academy of Oral and Maxillofacial Radiology and the American Academy of Orofacial Pain.

This position statement was developed by an ad hoc committee of the American Academy of Oral and Maxillofacial Radiology and the American Academy of Orofacial Pain. The committee reviewed pertinent literature and drafted recommendations for imaging. The statement provides evidence-based recommendations and clinical guidance to apply appropriate diagnostic imaging to evaluate the temporomandibular joint.

Assessment of aflatoxin B1-lysine adduct in children and its effect on child growth in Lahore, Pakistan.

Aflatoxin B1 is an important toxic food contaminant and there is very little information available about its exposure and effects on the health of the Pakistani population. Therefore, children (n = 238) aged 1-11 years were recruited in this study to estimate the levels of aflatoxin B1-lysine adduct and to measure its adverse effects on growth. Blood samples were analyzed to detect AFB1-lysine adducts through high-performance liquid chromatography. Socio-demographic information and anthropometry measurements were also obtained. All participants had detectable levels of AFB1-lysine adduct with a median concentration of 10.66 pg/mg albumin (95% CI: 8.6-12.4). Differences in area of residence (p < 0.05) and the father's employment (p < 0.05) were significant predictors for aflatoxin concentration levels in ordinary least square and quantile regression models (residence in 75th quantile and father employment in 90th quantile). Children aged from 5 to 11 years in the 5th and 90th quantiles of the regression model had a significant association with aflatoxin levels. A very high (50.4%, 120/238) prevalence of growth impairment (stunting, wasting, and underweight) was also observed in this study. Although we couldn't establish the effect of aflatoxin on growth impairment, children with low serum albumin levels (OR = 0.18; 95% CI: 0.05-0.56; p = 0.004) were likely to be at risk of wasting. Also, low birth weight was strongly associated with wasting (OR = 3.11; 95% CI: 1.36-7.03; p = 0.006) and underweight (OR = 4.60; 95% CI: 2.21-10.05; p= <0.001), while the mother's school level education had a correlation with child stunting (OR = 1.84; 95% CI: 1.07-3.22; p = 0.029). The high prevalence of growth impairment and high concentration of serum AFB1-lysine adduct levels in study participants demand immediate efforts to mitigate the adverse health outcomes in children in Pakistan.

Corrigendum: Disease specific symptoms indices in patients with celiac disease-A hardly recognised entity.

[This corrects the article DOI: 10.3389/fnut.2022.944449.].

Disease specific symptoms indices in patients with celiac disease-A hardly recognised entity.

Background: Celiac disease (CD) was considered a rare disease before and was perceivably only limited to children but now affects almost 1-2% of the global population. This abrupt increase in prevalence is due to advancements in diagnostic criteria and medical facilities but still many countries lack the basic data that can assess the severity of this health issue. The present study was conducted with the aim to assess the common but rarely diagnosed condition with the identification of its underlying secondary ailments. Materials and methods: Patients visiting public sector hospitals were recruited and tested for clinical symptoms secondary to gluten-containing foods (wheat and barley, etc.), followed by serological testing for immunoglobulin A, tissue transglutaminase A, and anti-endomysial antibodies. Only seropositive candidates were included in the endoscopic and biopsy examination for the features of villous atrophy and intestinal cell damage. The secondary ailments including anemia, growth retardation, and gastrointestinal symptoms were also documented for the tested positive patients. The modified European Society of Pediatric Gastroenterology, Hepatology, and Nutrition (ESPGHAN) criterion was followed throughout the study. Results: From 647 suspected cases from March 2018 to July 2019, 113 were confirmed with CD while 58% were female children and 42% were male children. The majority of them were from a lower class (75%) and 26% of them had a positive family history of CD. A total of 67% of patients with CD were underweight while wasting was observed in 38%, and 80% were stunted as well. Of the positively tested patients with CD, 49% had moderate anemia with 15% having severe anemia. Approximately 33% had hypoalbuminemia as well. The majority of them had a mild to severe range of gastrointestinal symptoms, such as abdominal pain, diarrhea, flatus, eructation, diarrhea, and steatorrhea. Conclusion: The study finding indicates an increased number of patients diagnosed with CD with an excessive sum of secondary ailments, such as anemia, growth failure, growth retardation, malnutrition, and gastrointestinal symptoms.

Guidelines for oral and maxillofacial imaging: COVID-19 considerations.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus causing the current coronavirus disease 2019 (COVID-19) pandemic, is not only highly infectious but can induce serious outcomes in vulnerable individuals including dental patients and dental health care personnel (DHCPs). Responses to COVID-19 have been published by the Centers for Disease Control and Prevention and the American Dental Association, but a more specific response is required for the safe practice of oral and maxillofacial radiology. We aim to review the current knowledge of how the disease threatens patients and DHCPs and how to determine which patients are likely to be SARS-CoV-2 infected; consider how the use of personal protective equipment and infection control measures based on current best practices and science can reduce the risk of disease transmission during radiologic procedures; and examine how intraoral radiography, with its potentially greater risk of spreading the disease, might be replaced by extraoral radiographic techniques for certain diagnostic tasks. This is complemented by a flowchart that can be displayed in all dental offices.

Molecular Fingerprinting and Hybridity Authentication in Cowpea Using Single Nucleotide Polymorphism Based Kompetitive Allele-Specific PCR Assay.

Optimization of a breeding program for increased genetic gain requires quality assurance (QA) and quality control (QC) at key phases of the breeding process. One vital phase in a breeding program that requires QC and QA is the choice of parents and successful hybridizations to combine parental attributes and create variations. The objective of this study was to determine parental diversity and confirm hybridity of cowpea F1 progenies using KASP (Kompetitive Allele-Specific PCR)-based single nucleotide polymorphism (SNP) markers. A total of 1,436 F1 plants were derived from crossing 220 cowpea breeding lines and landraces to 2 elite sister lines IT99K-573-1-1 and IT99K-573-2-1 as male parents, constituting 225 cross combinations. The progenies and the parents were genotyped with 17 QC SNP markers via high-throughput KASP genotyping assay. The QC markers differentiated the parents with mean efficiency of 37.90% and a range of 3.4-82.8%, revealing unique fingerprints of the parents. Neighbor-Joining cladogram divided the 222 parents into 3 clusters. Genetic distances between parents ranged from 0 to 3.74 with a mean of 2.41. Principal component analysis (PCA) depicted a considerable overlap between parents and F1 progenies with more scatters among parents than the F1s. The differentiation among parents and F1s was best contributed to by 82% of the markers. As expected, parents and F1s showed a significant contrast in proportion of heterozygous individuals, with mean values of 0.02 and 0.32, respectively. KASP markers detected true hybridity with 100% success rate in 72% of the populations. Overall, 79% of the putative F1 plants were true hybrids, 14% were selfed plants, and 7% were undetermined due to missing data and lack of marker polymorphism between parents. The study demonstrated an effective application of KASP-based SNP assay in fingerprinting, confirmation of hybridity, and early detection of false F1 plants. The results further uncovered the need to deploy markers as a QC step in a breeding program.

Genetic worth of multiple sets of cowpea breeding lines destined for advanced yield testing.

The objective of this study was to determine genetic potentials in eight sets of cowpea lines for grain yield (GY), hundred seed weight (HSDWT) and days to 50% flowering (DT50FL). A total of 614 F6 genotypes constituting the sets, grouped by maturity, were evaluated across two locations in Northern Nigeria, in an alpha lattice design, two replications each. Data were recorded on GY, HSDWT and DT50FL.Variance components, genotypic coefficient of variation (GCV), and genetic advance (GA) were used to decode the magnitude of genetic variance within and among sets. Genetic usefulness (Up) which depends on mean and variance to score the genetic merits in historically bi-parental populations was applied to groups of breeding lines with mixed parentage. Principal component analysis (PCA) was used to depict contribution of traits to observed variations. GY and DT50FL explained the variance within and between sets respectively. Genotypes were significantly different, although genotype-by-location and set-by-location interaction effects were also prominent. Genetic variance (δ2 G) and GCV were high for GY in Prelim2 (δ2 G = 45,897; GCV = 19.58%), HSDWT in Prelim11 (δ2 G = 7.137; GCV = 17.07%) and DT50F in Prelim5 (δ2 G = 4.54; GCV = 4.4%). Heritability varied among sets for GY (H = 0.21 to 0.57), HSDWT (H = 0.76 to 0.93) and DT50FL (H = 0.20 to 0.81). GA and percentage GA (GAPM) were high for GY in Prelim2 (GAPM = 24.59%; GA = 269.05Kg/ha), HSDWT in Prelim11 (GAPM = 28.54%; GA = 4.47 g), and DT50F in Prelim10 (GAPM = 6.49%; GA = 3.01 days). These sets also registered high values of genetic usefulness, suggesting potential application in non-full sib populations. These approaches can be used during preliminary performance tests to reinforce decisions in extracting promising lines and choose among defined groups of lines. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s10681-020-02763-y).

Seroprevalence and Associated Risk Factors of Bovine Brucellosis in District Gujranwala, Punjab, Pakistan.

Bovine brucellosis is a contagious zoonotic disease that causes economic losses through abortion and infertility. A cross-sectional study was designed to determine the seroprevalence and associated risk factors of bovine brucellosis in district Gujranwala of Punjab, Pakistan. A total of 220 bovine sera (112 from buffaloes, 108 from cattle) from 46 unvaccinated herds were collected. Parallel testing by the Rose Bengal Plate Test (RBPT) and Indirect Enzyme-linked Immunosorbent Assay (I-ELISA) showed a 58.7% (27/46) herd-level and 22.7% (50/220) animal-level seroprevalence. Seroprevalence was higher (p < 0.001, OR = 7.62) in adult animals (37.2%) compared to younger animals (4.9%). A herd size of >10 animals (p = 0.021, OR = 7.83), less housing space (p = 0.037, OR = 6.39) and history of abortion at the farm (p = 0.023, OR = 5.6) were found as risk factors associated with the seropositivity of brucellosis. There was a substantial agreement between the RBPT and I-ELISA results (Cohen's kappa coefficient (κ) = 64.16, percent agreement = 89.5%). In conclusion, a relatively higher seroprevalence was found compared to the previous reports from the country. Standardization and validation of the advanced diagnostic tests would be needed. Biosecurity, personal protection, quarantine measures and routine screening of animals at the farm level and disease awareness programs and consumption of pasteurized milk in the human population will be helpful in preventing the transmission/zoonosis of the disease.

Prevalence and molecular diagnosis of Trypanosoma evansi in Nili-Ravi buffalo (Bubalus bubalis) in different districts of Punjab (Pakistan).

The prevalence of Trypanosoma evansi was investigated in 1,250 Nili-Ravi buffaloes of mixed age and sex by polymerase chain reaction (PCR) for the first time in Pakistan. DNA of the trypanosomes was isolated with TRIREAGENT®. The assay was employed using primers ESAG 6/7, specific for a 237-bp fragment from T. evansi genomic DNA. The samples were screened for the presence of T. evansi also by stained thin smear. Forty-four (3.5%) samples were positive by microscopy, while 97 (7.7%) samples were identified by PCR, indicating the high sensitivity of PCR for surveying the disease in epidemiological studies.