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Raising small ruminants is the main source of income for farmers in Pakistan especially in rural areas of Dera Ghazi Khan in Punjab. Despite having large sheep population, the prevalence of intra-erythrocytic protozoa, Theileria (T.) lestoquardi, has never been reported from this area. This study was conducted to fill this knowledge gap and 333 blood samples of apparently healthy small ruminants (168 sheep and 165 goats) along with their epidemiological data were collected from Dera Ghazi Khan district during August till November 2022. The polymerase chain reaction (PCR) analysis amplified a 785 base pair amplicon specific for the Merozoite surface antigen (ms 1–2) gene of T. lestoquardi in 2 out of the 168 (3.3%) sheep blood samples, while no goat blood sample out of 165 was found to be infected with T. lestoquardi. DNA sequencing confirmed the presence of Theileria lestoquardi in both samples and phylogenetic analysis revealed that these amplicon resembled the partial ms 1–2 gene sequences detected in small ruminants from Pakistan, India Iran and Egypt. All the studied epidemiological factors (age, sex, breed, size of herd, dogs with herd, composition of herd, size of herd and Tick burden on sheep) were not found associated with the prevalence of T. lestoquardi. In conclusion, this study reports a low prevalence of T. lestoquardi infection in the Dera Ghazi Khan District of Punjab, Pakistan. The data generated from this work will help pave the way for the prophylactic detection and control of ovine and caprine theileriosis in the region. [ABSTRACT FROM AUTHOR]

Objective: To compare the frequency and types of anemia in subclinical hypothyroid and euthyroid patients. Study Design: Comparative cross sectional study. Place and Duration: Department of Medicine, PNS Shifa, Karachi Pakistan, from Jun 2019 to Dec 2019. Methodology: All patients who fulfilled the inclusion criteria and visited Medicine Department, PNS Shifa Karachi were included in the study. After ethical approval and informed written consent patients were divided into two groups, subclinical hypothyroidism group (149 patients) and euthyroid group (148 patients). Blood samples of all included patients were sent for hematocrit, mean cell volume and hemoglobin level to assess the outcome i-e anemia & its different types. Results: A total of 297 patients were included in the study. Amongst the sub clinically hypothyroid patients, anemia was noted in 40 patients (26.9%), while in the euthyroid group, it was noted in 23 patients (15.5%). In the subclinical hypothyroidism group, the type of anemia was microcytic in 13 patients (8.7%), macrocytic in 7 patients (4.7%) and normocytic in 20 patients (13.4%); while in the euthyroid group, anemia was microcytic in 7 patients (4.7%), macrocytic in 4 patients (2.7%) and normocytic in 12 patients (8.1%). Conclusion: The commonest type of anemia found in sub clinically hypothyroid patients is normocytic normochromic anemia. The frequency of anemia was 26.9% in the subclinical hypothyroid group and 15.5% in the euthyroid group. [ABSTRACT FROM AUTHOR]

Theileria annulata (T. annulata) and Anaplasma marginale (A. marginale) are among the most extensively reported tick borne pathogens and are associated with huge economic losses worldwide. A total of 298 cattle blood samples were screened to report the presence of these two pathogens. The samples were collected from apparently healthy cattle (Achai, n = 155, Jersy, n = 88 and crossbred, n = 55) in Bajaur district of Khyber Pakhtunkhwa (KPK) during June and July of 2022. A total of 31 out of 298 cattle (10.4%) were found infected with T. annulata as PCR amplified a 156 base pair fragment from Tams-1 gene of T. annulata from their blood. While 16/298 animals (5.4%) were found infected with A. marginale as they amplified a 382 base pair fragment specific for msp5 gene of this bacterium. Three animals (1%) were found co infected. Cattle susceptibility to T. annulata infection was significantly higher than A. marginale infection (P < 0.001). Phylogenetic analysis revealed that Pakistani isolates of both detected pathogen clustered together and were closely related isolates from worldwide countries. Prevalence of T. annulata varied significantly among the sampling sites (P = 0.05) while no such association was observed for A. marginale among the tested cattle. Epidemiological data analysis revealed that none of the studied risk factors was found associated either with the prevalence of T. annulata or A. marginale (P > 0.05) among enrolled cattle. In conclusion, our study has revealed a relatively higher prevalence of T. annulata than A. marginale in cattle from the Bajaur district in KPK. This information is important for improving the productivity of the livestock sector, which is one of the main sources of income in the country. It is recommended that this data be taken into account for the development and implementation of effective tick control programs, as well as for the improvement of livestock management practices to prevent and manage TBDs in Pakistan. [ABSTRACT FROM AUTHOR]

The mosquito-borne virus that causes dengue fever has emerged as a major public health problem in recent decades. Dengue fever prevalence in District Rawalpindi was investigated in this study, between August to October, 2022. About 160 blood samples were taken from patients at the Islamabad Medical Lab (Rawalpindi, Punjab) who were suspected of having dengue fever based on their symptoms. Dengue IgM, IgG, and NS1 (Nonstructural protein antigen of virus) serology tests were carried out. About 116 males and 44 females were found to have contracted dengue fever. The 21-30 age group was the largest single demographic, 78 people from Rawalpindi, 47 people from Taxila and 35 people from Kahuta were the worst hit in the district Rawalpindi. As a result, the health department needs to take charge and start spreading awareness about the importance of keeping public spaces clean. Efforts to combat diseases and infections spread by vectors should actively involve the local populace. [ABSTRACT FROM AUTHOR]

Objective: To determine the frequency of Blood Group A2 genotype among Group A Pakistani whole blood donors. Study Design: Cross-sectional study. Place and Duration of Study: Armed Forces Institute of Transfusion (AFIT), Rawalpindi Pakistan, from Jan 2019 to Jan 2020. Methodology: One thousand (1000) healthy and unrelated blood donors were selected. The blood samples were typed for ABO, and those of Blood Groups-A and AB were further subtyped with the help of anti-A1 lectin to categorize them as A1, non-A1, A1B and non-A1B Groups. Next, DNA of non-A1 samples was extracted, and a Polymerase Chain Reaction using sequence-specific primers (PCR-SSP) for type A2 was performed, followed by polyacrylamide gel electrophoresis (PAGE). Results: Among one thousand blood donors, 247(24.7%) were typed as Blood Group-A, 94(9.4%) as AB-Group, 339(33.9%) B-Group and 320(32%) O Group with variable strength of reaction with ABO antisera. A and AB Blood Groups were further subgrouped as A1 202(20.2%), A1B 77(7.7%), non-A1 45(4.5%) and non-A1B 17(1.7%). Anti-A1 antibodies were detected in 6(13.3%) of non-A1 samples. PCR of non-A1 samples showed 32(12.9%) to be genotypically A2, and the remaining 13(5.2%) were not A2 and were not further resolved. Conclusion: Blood Group A2 is not a rare Subgroup in our population. PCR-SSP is a more specific technique than anti-A1 lectin for establishing the Blood Group status of an individual. [ABSTRACT FROM AUTHOR]

Objective: To determine the prevalence of A1 and A2 subgroups of blood group A in healthy blood donors of northern Pakistan. Study Design: Cross sectional study. Place and Duration of Study: Armed Forces Institute of Transfusion, Rawalpindi Pakistan, from Sep to Dec 2017. Methodology: The blood samples of donors reported during the study period at Armed Forces Institute of Transfusion were collected. ABO typing was performed and those in group A and AB were further subtyped using anti-A1 lectin and thus the individuals were labelled as A1 (and A1B) or A2 (and A2B). The serum of individuals with groups A2 and A2B was tested for anti-A1 based on the reaction with A1 red cells. Results: Out of total 4485 donors, 1116 (24.88%) were of blood group A and 422 (9.41%) individuals were of AB group. Out of 146 (13.08%) individuals among 1116 blood group A donors were of subgroup A2. While out of 422 of blood group AB, 80 (18.95%) were A2B. 21 (14.38%) out of 146 individuals of A2 blood group, had anti-A1 antibody while among 80 individuals with A2B blood group only 2 (2.50%) had anti-A1 antibody in their serum. Conclusion: Blood group A2 is quite prevalent in our population. Although anti-A1 is present in significant fraction of A2 blood group individuals, the routine testing is not needed to identify the A2 group and cross match compatibility is the only requirement as anti-A1 is rarely active at 37° C. [ABSTRACT FROM AUTHOR]

The purpose of this study was to identify the seroprevalence of Toxoplasma gondii (T. gondii) and hematological alterations in captive pheasant species of Bahawalpur Zoo, Bahawalpur, Pakistan. The blood samples of 100 birds belonging to three different species viz. ring-necked pheasant (Phasianus colchicus) (n=46), green pheasant (Phasianus versicolor) (n=40), and silver pheasant (Lophura nycthemera) (n=14) were analyzed through Latex Agglutination Test (LAT) and Enzyme Linked Immunosorbent Assay (ELISA). Seroprevalance through LAT in ring-necked, silver and green pheasants was 10.86%, 7.14% and 7.5%, and through ELISA was 32.60%, 22.5% and 14.28%, respectively. Adults and males had higher prevalence as compared to their counterparts, though, statistically nn-significant (P=0.05). In all the studied species, hematological results revealed that mean values of Hb and TEC were higher (P=0.05) in noninfected birds as compared to infected ones. This is a preliminary study of a kind and needs further research with large number of sample and population across the country. [ABSTRACT FROM AUTHOR]

Objective: The current study aimed to investigate the ABO and rhesus (Rh) blood group frequency in the people of District Faisalabad and Sheikhupura, Punjab Province, Pakistan. The retrospective study was conducted on more than thirty thousand people including both male and female patients admitted to the Tehsil Headquarter Hospital, Safdarabad and The Best Hospital, Faisalabad. Blood samples were taken from each subject and subsequently ABO and Rh blood groups were evaluated separately. The antigen antibody agglutination slide test for blood grouping (ABO) and Rh were used to assess the blood group frequencies. Results: The frequencies of ABO blood group distribution indicated that blood group B was predominant in the people of Safdarabad followed by O, A and AB respectively. While, among people of Faisalabad, blood group O was predominant followed B, A and AB respectively. Rh negative phenotype was found lesser distributed as compared to the positive Rh phenotype. [ABSTRACT FROM AUTHOR]

Anaplasmosis is a tick-borne disease caused by obligate intercellular gram-negative bacteria, Anaplasma (A.) marginale. The present study reports on seasonal prevalence, epidemiology, and phylogeny of A. marginale in three cattle breeds from District Layyah, Southern Punjab, Pakistan. A total of 844 blood samples (Cross = 300, Holstein Friesian = 244, Sahiwal breed = 300) from apparently healthy cattle on seasonal basis were collected along with epidemiological data during May 2018 till April 2019. Polymerase chain reaction generated 265 base-pair amplicon specific for major surface protein-1b encoding gene of A. marginale in 8.6% (73/844) of enrolled cattle. The highest prevalence was observed during autumn (18.3%) followed by summer (9.7%) and winter season (7.1%). Holstein Friesian breed was most susceptible to A. marginale infection (13.1%) followed by Sahiwal (7.6%) and cross breed (6%). Representative amplified partial gene sequences of A. marginale were submitted to GenBank (Accession numbers MK032842 and MK032843). 37/844 (4.3%) Giemsa-stained blood smears were found positive for Anaplasma spp. Small number of ticks including Hyalomma anatolicum, Hyalomma excavatum, Rhipicephalus microplus, Haemaphysalis punctata were identified from cattle but none of them was found PCR positive for the presence of A. marginale. Analysis of epidemiological factors revealed that female cattle and farm with water supply from pool, farms where other dairy animals and dogs were living with cattle and dogs having ticks load on them had significant association with A. marginale prevalence. It was observed that white blood cell, lymphocytes (%), monocytes (%) hematocrit, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration were significantly disturbed in A. marginale-positive than negative cattle. [ABSTRACT FROM AUTHOR]

Surra caused by Trypanosoma evansi is a well-known infection in camel and equines and to a lesser extent in other domestic and wild animals in Pakistan. The purpose of this study was to evaluate the burden and the precise cause of surra in dogs. A total of 160 blood samples were collected, and 3.75% (6/160) were found positive by microscopic examination and 2.5% (4/160) by PCR. One female (25%) and 3 male (75%) dogs were found positive by PCR. Among them, the 2 positive dogs were from Pakistani bully breed and the other 2 from local/non-descriptive breeds. All 4 dogs were between 2-4 years of age. A more precise study is needed for determination of the associated risk factors with this infection. To the best of our knowledge, this is the first-ever report of specific Trypanosoma evansi detection in dogs in Pakistan. [ABSTRACT FROM AUTHOR]

Aim: The present study was designed to check the molecular detection of Anaplasma marginale and Theileria annulata in blood samples of horses and donkeys collected from Dera Ghazi Khan District in Punjab and to document their phylogenetic origin and their association with studied epidemiological factors (sex and age) and complete blood count parameters, if any. Methods and results: A total of 195 blood samples were collected from apparently healthy horses (N = 141) and donkeys (N = 54). A. marginale DNA was detected by PCR in 4.9% (7/141) horse and in 9.2% (5/54) of donkey blood samples. Prevalence of T. annulata was 5.6% (8/141) and 11.1% (6/54) in horse and donkey samples, respectively. While 1.4% (N = 2) horses and 3.7% (N = 2) donkeys were found co-infected with both parasites. Representative amplicon for both parasites was confirmed by DNA sequenced and partial DNA sequence of the major surface protein–1b encoding gene of A. marginale and cytochrome b gene from T. annulata were submitted to the GenBank database under the accession number MK792344-MK792348. Epidemiological data analysis revealed that female horses were more prone to A. marginale (P = 0.02) while female donkeys were more susceptible to A. marginale (P < 0.001) and T. annulata (P < 0.001) infection. It was observed that horse and donkey infected either with Anaplasma marginale or Theileria annulata had significantly disturbed red and white blood cell counts and their associated parameters. Conclusion: This is a first ever study regarding molecular detection of A. marginale and T. annulata in equine blood samples from Pakistan. We recommend that this multiplex PCR protocol should be used for the detection of Anaplasma marginale and Theileria annulata in livestock for their proper diagnosis and treatment. [ABSTRACT FROM AUTHOR]

It is a common perception among dairy farmers in Pakistan that oxytocin (OT) injection negatively effects on buffalo reproduction parameters. Therefore, current study was conducted to investigate the outcomes of OT on numbers of AI per conception, pregnancy rate, embryonic and fetal losses (EFL). For this purpose, Nili-Ravi buffaloes (n=23) were randomly divided into three groups, G1; n=7 (control), G2; n=8 (low dose 10 i.u.) and G3; n=8 (high dose 30 i.u.). The OT injection was given twice daily at morning and evening milking until 154 days post calving. At heat, AI was performed. Ultrasonography and blood sampling were done on days 30, 45 and 60 post AI to monitor pregnancy and progesterone, respectively. Results showed significant (P=0.05) increase of AI services per conception in G2 & G3 as compared to G1. Furthermore, results revealed that chances of the animals to become pregnant were 0.227 and 0.094 times in low (G2) and high doses (G3) in comparison to control (G1), respectively, and was lower significantly (P=0.05). Similarly, pregnancy rate was remained unaffected and was non-significant (P>0.05) on day 45 vs day 30. However, it was significantly (P=0.05) lower on day 60 vs day 30. Overall, pregnancy rate was decreased (P=0.05) in oxytocin treatments comparable to control. Parity and days in milk had no significant effect on pregnancy rate among treatments (G1, G2 and G3), respectively. Moreover, EFL losses were non-significant (P>0.05) among treatments but losses trend was more in G2 & G3 parallel to G1. Result also showed non-significant (P>0.05) decrease in P4 on days 30 and 45 while significant (P=0.05) on day 60 in G2 & G3 as compared to G1 among treatment groups. It may be concluded that long term OT injections of both low and high doses cause significant losses in terms of increase in AI services per conception and decrease pregnancy rate with higher EFL in Nili-Ravi buffaloes. [ABSTRACT FROM AUTHOR]

Main focus is to study the three districts of Punjab as well as the nearby areas where Kajli breed exists. DNA quality and quantity was tested by 1% agarose gel. DNA and amino acid content was also analyzed from the blood samples. Majority of amino acids of different breeds are similar to each other which showed that their origin is similar from Urial. However, Alanine content in sheep located from Sargodha was higher than those located with other areas. . It is suggested that the minute difference in these breeds is due to different kinds of breeding that is done in different areas of Pakistan. Sequence of the cytochrome b gene was visualized by the use of Chromas software. Position of band in similar region showed that all the breed belongs to similar origin. Populations of Mianwali, Khushab and Sargodha regions are clustered into one group. The cluster patterns were concordant with their geographic localities [ABSTRACT FROM AUTHOR]

Objectives: To determine frequencies of ABO blood group in T2DM in tertiary Care, Pakistan. Study Design: Descriptive, Cross-sectional study. Place: Conducted in the department of medicine, Ghulam Muhammad Mahar Medical Teaching Hospital (GMMMC). Duration: From January 2016 to July 2016. Methods: With consent of patients having T2DM of both genders and 30-70 years of age. Non-probability, Consecutive sampling was used. Total of 196 patients were enrolled during this time period. After taking consent, blood samples taken from peripheral vein for blood grouping were sent to the Central Laboratory GMMMC. Laboratory technician determined blood group using standard techniques. All this information was collected through a self-structured Performa. Results: Participants in our study were from age range 30 to 70 years with mean age of 53.26 ± 7.28 years. Out of the 196 patients, 110 (56.1%) were female and 86 (43.9%) were male. The prevalence of blood group A, B, AB and O in patients with ischemic stroke was found to be as of following; 34 patients having blood group A (17.3%), 74 patients having blood group B (37.8%), 18 patients having blood group AB (9.2%) and 70 patients having blood group O (35.7%) respectively. Stratification was also done on gender basis. 75.5% (n=148) had positive family history. Conclusion: Blood group B +ve was the most prevalent blood group in T2DM in our study. [ABSTRACT FROM AUTHOR]

Background & Objective: Errors in the pre-analytical phase constitute about 68% of all lab diagnostic errors and they occur mostly during specimen collection phase. The aim of our study was to assess awareness of resident and internee doctors regarding pre-analytical errors during venous blood sampling and handling. Methodology: This cross-sectional questionnaire-based study was conducted at Mayo Hospital, Lahore. One hundred and eighty residents and internees were included, whereas doctors working solely in Emergency & Pathology departments were excluded. Data were collected through questionnaire consisting of 29 questions. Results: A mean ± SD of correct responses was 10.61 ± 3 was found in total sample of 180 participants out of 25 selected questions. Higher mean values were observed in residents as compared to internees, in Pediatrics subjects as compared to other specialties and in graduates from outside Pakistan, as compared to local graduates. Amongst the questions with the lowest correct answer ratio were: forceful pulling of plunger of syringe (36.1%), awareness regarding evacuated tubes system (30%), vials used for hematological tests (5.6%), glucose estimation tests (4.4%) and plasma electrolytesestimation (0.0%). Conclusion: Study shows a lack of awareness of stages of pre-analytical phase, and thus of possible errors in this phase. This directly influences quality of lab results, compromising the time, finances and health of patients. Therefore, the situation demands prompt correction. [ABSTRACT FROM AUTHOR]

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Objective: To determine median HbA1c level in patients screened for DM and compare the diagnostic accuracy of HbA1c with the currently recommended ADA cut off, using 2 hours (h) plasma glucose (2-h PG) post 75 grams oral glucose tolerance test (OGTT) as gold standard. Study Design: Cross sectional study. Place and Duration of Study: Aga Khan University Hospital Karachi, from Jul 2014 to Nov 2015. Patients and Methods: Consecutive subjects screened by OGTT for diabetes mellitus (DM) were included. Blood sample for plasma glucose and HbA1c were analyzed on ADVIA 1800. Results: Total 146 subjects were included with median age 45 (IQR 54.2-35) years; 53% (n=77) being females. Median HbA1c of the study subjects was 6% (IQR 6.6-5.6). Positive correlation was observed between age and HbA1c [5.7% (IQR 6.2-5.3) <40 y, 6.2% (IQR 6.8-5.8) in >40 y subjects; r 0.34, p=0.000). Males had higher HbA1c than females [6.1% (IQR 6.8-5.7) years. 5.9% (IQR 6.4-5.4); r 0.17, p=0.036). HbA1c levels were significantly different amongst the different ethnic groups residing in Pakistan (p-value<0.03). HbA1c was positively correlated with FPG (r=0.59, p-value<0.001) and 2-h PG (r=0.56, p-value<0.001). Sensitivity, specificity, PPV, NPV for HbA1c at cutoff of 6.5% in diagnosing diabetes mellitus (DM) were 70%, 89%, 72% and 80% respectively. Conclusion: High median HbA1c levels is noted in our subjects. At cut off level of HbA1c ≥6.5%, 70% of subjects with DM were diagnosed using ADA criteria of 2-h PG for diabetes as gold standard. Ethnic differences were also observed in levels of HbA1c. There is a need to define cut off for our population. [ABSTRACT FROM AUTHOR]

The study was aimed to isolate BoHV-1 circulating in Lahore, Pakistan and its genetic characterization. For this purpose, blood samples were collected from different areas of district Lahore through convenient sampling technique. Out of 100 blood samples, 69 (69%, 95% CI: 58.86-77.66) samples have shown seropositivity against BoHV-1 through ELISA. For the isolation of BoHV-1, nasal swab samples were collected from 69 seropositive cattle and buffalo. Out of 69 nasal swab samples, only 6 samples have shown the visible cytopathic effect and confirmed through nested polymerase chain reaction by targeting glycoprotein B (gB). The phylogenetic analysis showed that local isolates showed similarity to subtype 1.1 of BoHV-1 and had 99-100% homology with Cooper strain. It was concluded that BoHV-1.1 is being circulating in Pakistan and further studies are needed which will help for clear understanding of virus characterization and development of effective local vaccine. [ABSTRACT FROM AUTHOR]

Background: Individual, comprehensive, and written informed consent is broadly considered an ethical obligation in research involving the sampling of human material. In developing countries, however, local conditions, such as widespread illiteracy, low levels of education, and hierarchical social structures, complicate compliance with these standards. As a result, researchers may modify the consent process to secure participation. To evaluate the ethical status of such modified consent strategies it is necessary to assess the extent to which local practices accord with the values underlying informed consent. Methods: Over a 2-week period in April 2014 we conducted semistructured interviews with researchers from a genetic research institute in rural Pakistan and families who had given blood samples for their research. Interviews with researchers focused on the institute’s requirements for consent, and the researchers’ strategies for and experiences with obtaining consent in the field. Interviews with donors focused on their motivation for donating samples, their experience of consent and donation, and what factors were central in their decisions to give consent. Results: Researchers often reported modifications to consent procedures suited to the local context, standardly employing oral and elder consent, and tailoring information to the social education level of donor families. Central themes in donors’ accounts of their decision to consent were the hope of getting something out of their participation and their remarkably high levels of trust in the researchers. Several donor accounts indicated a degree of confusion about participation and diagnosis, resulting in misconceived expectations of therapeutic benefits. Conclusions: We argue that while building and maintaining trusting relationships in research is important—not least in developing countries—strategies that serve this endeavor should be supplemented with efforts to ensure proper provision and understanding of relevant information, specifically about the nature of research and measures for individual consent and opt-out. [ABSTRACT FROM AUTHOR]

Leptospirosis is a worldwide emerging infectious disease of zoonotic importance and large epidemics and epizootics have been reported all over the globe. A cross survey study was conducted to estimate seroprevalence of human leptospirosis in climatically distinct regions of Pakistan and to identify the risk factors associated with the disease. Blood samples from 360 humans were collected through convenient sampling, 120 from each of three study areas. Serological testing was performed using ELISA kit as per manufacturer’s recommendations. The results showed an overall prevalence of 40.83% (95% CI; 35.71–46.11). Statistical analysis showed significant (P < .05) differences in leptospiral seroprevalence in three different geographic locations, with highest in humid sub-tropical climatic region (50.83%; 95% CI; 41.55–60.07), followed by semi-arid region (44.16%; 95% CI; 35.11–53.52) and lowest in hot and dry region (27.50%; 95% CI; 19.75–36.40). After multivariate analysis age, gender, exposure to flooding water, source of water usage, disinfection schedule of surroundings and history of cut and wound were found significantly associated with the seropositivity of Leptospira . The present study, first to uncover seroprevalence of human Leptospira in different climatic regions of Pakistan, alarms about effect of climate on prevalence of Leptospira in the region. [ABSTRACT FROM AUTHOR]

The bovine leptospirosis is an economically important zoonotic disease of flood affected areas worldwide, but scarce information is available about its epidemiology in Pakistan. This is a first study on sero-epidemiology of bovine leptospirosis in Pakistan. The objectives of this study were to investigate the sero-prevalence and associated risk factors of bovine leptospirosis in flood affected zone of Punjab, Pakistan. A total of 385 serum samples were randomly collected from four tehsils of district Muzaffargarh, Pakistan. The serum samples were subjected to indirect ELISA for the detection of anti-leptospira antibodies. The overall sero-prevalence of leptospirosis was 30.39%. The prevalence was significantly higher ( p < 0.001) in cattle (56.25%) than buffaloes (4.66%). The key risk factors identified based on multivariable logistic regression were; confined system of rearing, flooded area, and lesser to graduate level of education as risk factors for leptospirosis. The values of hematological parameters varied significantly ( p < 0.05) for Hb, TEC and PCV while there was no significant ( p > 0.05) difference among TLC values among sero-positive and sero-negative animals. The serum biochemical profile revealed significant differences ( p < 0.05) in values of ALT, AST and creatinine among the sero-positive and negative animals. Hence, it can be concluded that leptospirosis is an emerging and neglected disease in flood affected zone of Pakistan, and the disease needs to be explored comprehensively in other parts of the country to sort out solid strategies for its control and eradication. [ABSTRACT FROM AUTHOR]

Background: Dengue virus is the causative agent of dengue fever, a vector borne infection which causes selflimiting to life threatening disease in humans. A sero-epidemiological study was conducted to understand the current epidemiology of dengue virus in Pakistan which is now known as a dengue endemic country after its first reported outbreak in 1994. Methods: To investigate the prevalence of dengue virus in Pakistan during 2009-2014, a total of 9,493 blood samples were screened for the detection of anti-dengue IgM antibodies using ELISA. Clinical and demographic features available with hospital records were reviewed to ascertain mortalities related to dengue hemorrhagic shock syndrome. Results: Out of 9,493 samples tested, 37% (3,504) were found positive for anti-dengue IgM antibodies. Of the seropositive cases, 73.6% (2,578/3,504) were male and 26.4% (926/3,504) were female. The highest number (382/929; 41.1%) of sero-positive cases was observed among the individuals of age group 31-40 years. The highest number of symptomatic cases was reported in October (46%; 4,400/9,493), and the highest number of sero-positive cases among symptomatic cases was observed in November (45.7%; 806/1,764). Mean annual patient incidence (MAPI) during 2009-2014 in Pakistan remained 0.30 with the highest annual patient incidence (11.03) found in Islamabad. According to the available medical case record, 472 dengue related deaths were reported during 2009-2014. Conclusion: The data from earlier reports in Pakistan described the dengue virus incidence from limited areas of the country. Our findings are important considering the testing of clinical samples at a larger scale covering patients of vast geographical regions and warrants timely implementation of dengue vector surveillance and control programs. [ABSTRACT FROM AUTHOR]

From 2013 to 2015, the National Institute of Health, Pakistan, received 1,270 blood samples of suspected dengue cases reported from inpatient and outpatient departments of various hospitals in Khyber Pakhtunkhwa (KPK) province. In this study, we determined the circulating dengue virus (DENV) serotypes using real-time reverse transcriptase (RT)- PCR to understand the serotype-based epidemiology of DENV. All four serotypes (DENV-1 [6%], DENV-2 [33%], DENV-3 [47%], and DENV-4 [0.1%]) were found circulating during the study period. Our findings suggest the need for an active surveillance system coupled with the laboratory diagnosis, especially in the chronic endemic areas of the country. Public awareness programs are needed for effective control and prevention of outbreaks in the future. [ABSTRACT FROM AUTHOR]

Background/aim: The prevalences of Plasmodium falciparum and Plasmodium vivax are increasing rapidly in Pakistan, but recent data on the epidemiology of malaria are not properly reported with scarce diagnostic methods for quick diagnosis. This study was designed to determine the current prevalence and distribution of Plasmodium species in the vicinity of Rawalpindi and Islamabad and report on the validity of the immunochromatographic test (ICT) in diagnosing malarial infections. Materials and methods: A total of 1500 blood samples obtained from a local hospital were screened during the course of this study via microscopic examination and ICT. Results: It was seen that malaria was highly endemic in this region. Both P. vivax and P. falciparum were prevalent in all age groups with high seasonal variations, showing a summer peak for P. vivax and a winter peak for P. falciparum. In a comparative study of the diagnostic methods it was observed that the ICT is 95% sensitive and 100% specific for both P. falciparum and P. vivax, while microscopic study was 100% sensitive and 96.8% specific. Conclusion: Epidemiological study of the malarial parasites showed that majority of the patients were from Rawalpindi as compared to Islamabad and that P. vivax was the dominant cause of malarial infection. [ABSTRACT FROM AUTHOR]

Background: Fusion oncogenes (FOs) resulting from chromosomal abnormalities have an important role in leukemogenesis in pediatric B cell acute lymphoblastic leukemia (ALL). The most common FOs are BCR-ABL, MLL-AF4, ETV6-RUNX1, and TCF3-PBX1, all of which have important prognostic and drug selection implications. Moreover, frequencies of FOs have ethnic variations. We studied Pakistani frequencies of FOs, clinical pattern, and outcome in pediatric B-ALL. Methods: FOs were studied in 188 patients at diagnosis using reverse transcriptase-polymerase chain reaction (RT-PCR) and interphase fluorescent in situ hybridization (FISH). Data were analyzed using SPSS version 17 (SPSS Inc., Chicago, IL, USA). Results: FOs were detected in 87.2 % of patients. Mean overall survival was 70.9 weeks, 3-year survival was 31.9 %, and 3-year relapse-free survival was 18.1 %. Four patients died of drug toxicities. ETV6-RUNX1 (19.14 %) had better survival (110.9 weeks; p = 0.03); TCF3-PBX1 (2.1 %) was associated with inferior outcome and higher central nervous system (CNS) relapse risk; MLL-AF4 (18.1 %) was more common in the 8- to 15-year age group (24/34; p = 0.001) and was associated with organomegaly, low platelet count, and poor survival; and BCR-ABL (47.9 %) was associated with older age (7-15 years, 52/90), lower remission rates, shorter survival (43.73 ± 4.24 weeks) and higher white blood cell count. Overall, MLL-AF4 and BCR-ABL were detected in 66 % of B-ALL, presented in later childhood, and were associated with poor prognosis and inferior survival. Conclusions: This study reports the highest ethnic frequency of BCR-ABL FO in pediatric ALL, and is consistent with previous reports from our region. Poor prognosis BCR-ABL and MLL-AF4 was detected in two-thirds of pediatric B-ALL and is likely to be the reason for the already reported poor survival of childhood ALL in South-East Asia. Furthermore, MLL-AF4, usually most common in infants, presented in later childhood in most of the ALL patients, which was one of the unique findings in our study. The results presented here highlight the need for mandatory inclusion of molecular testing for pediatric ALL patients in clinical decision making, together with the incorporation of tyrosine kinase inhibitors, as well as hematopoietic stem cell transplantation facilities, to improve treatment outcome for patients in developing countries. [ABSTRACT FROM AUTHOR]

Asthma is a chronic inflammatory and remodeling disorder of the airways, in which many cells, cellular elements, and cytokines play important roles. The role of tumor necrosis factor-α (TNF-α) in asthma is unclear in Pakistani population. The aim of this study was to assess the relationship between TNF-α-308 polymorphism and asthma. Polymorphism of TNF-α (G-308-A locus; rs 1800629) in 329 asthmatic patients and 151 healthy controls was evaluated. DNA was prepared from blood samples of cases and controls. Samples were genotyped for TNF-α 308 G/A polymorphism. There was no significant difference in the frequency of GG (OR 1.049 with 95% CI 0.68-1.63) and GA (OR 0.987 with 95% CI 0.64-1.53) genotypes of TNF-α-308. The AA genotype was absent in cases and only one AA genotype was observed in the controls. The genetic polymorphism of TNF-α does not seem to be associated with asthma in Pakistani population. [ABSTRACT FROM AUTHOR]

Clinical signs, viz lethargy, increased heart rate and reduced appetite, making trypanosomiasis a possible differential diagnosis, were found in five out of twenty semi-captive Asiatic black bears (Ursus thibetanus) in a sanctuary, located in Kund, District Sawabi, KPK, Pakistan. Microscopic examination of blood samples of bears expressing clinical signs and symptoms revealed the presence of haemoflagellates, which was found to be trypanosomes. Subsequently, the PCR technique was exploited to screen for the presence of trypanosomal species in all bears' blood samples. Blood samples from 20 individual bears were screened using three sets of primers specific to Trypanosoma evansi species. Three primer pairs used are equally effective in successful detection of the parasite. Two out of five, diseased bears died prior to any trypanosoma specific medication while the rest were given an administered dose of Melarsomine (Immiticide). The treated bears survived and were assured to be aparasitemic on post-treatment examination after six weeks. [ABSTRACT FROM AUTHOR]

The man's existence is closely linked with animals in multiple use, to earn livelihood and even used as food. The fodder and forages are the major nutritional items along with other supplements. In Pakistan and sub-continent, the shortage of fodder and forages is very common particularly in kharif season because shortage of the fresh water resources. The waste water is manly passed down as supplemental irrigation source to irrigate field crops including fodder and forages. The studies showed the provisions of Iron (Fe) in feces, forages, soil, fodder and grazing animal's blood data collect from the Sargodha, Jhang, Layyah and Bhakkar districts of the Punjab. Five sites, three animal's (Cow, Sheep and Goat) and two origins (Blood and Feces) were selected. Standard samples preparation methods were worker to check occurrence of the heavy rock's metals in the forage and soil samples. The Atomic Absorption Spectrophotometer was used to check Iron (Fe). The results revealed that Fe were found in safe limit, as per WHO. [ABSTRACT FROM AUTHOR]

The present field study was conducted for confirmation of chicken infectious anemia (CIA) in White Leghorn (WL) commercial layer birds. A total of 60 farms were investigated. Birds from each farm ware necropsied for the presence of lesions on different visceral organs. Samples of blood and different tissues were collected for hematology, histopathology, DNA extraction and PCR amplification using specific primers for CIA virus. There was severe anemia indicated by low hematocrit values (10.9 to 17.2%) and hemoglobin concentration (5.3 to 6.7 g/dl). The petechial hemorrhages were present on subcutaneous tissue, epicardium, endocardium and gizzard mucosa. The liver and bone marrow were pale in appearance. The mortality ranged from 5 to 14 % on different farms. Samples of liver and spleen from 15 farms were subjected to PCR analysis for CIAV infection by amplifying the 186-bp region on highly conserved VP-2 coding gene using CAV1 and CAV2 primer pair. Presence of CIAV was confirmed in 67 and 33 percent samples of liver and spleen, respectively. A total of 13/15 farms (87%) were found positive for CIA. The results of present study confirmed the presence of CAV infection in WL commercial layer birds in current outbreak. It is concluded that extensive molecular epidemiological studies are required at national level to assess the prevalence of disease. Breeder flocks should be vaccinated to control CIA in commercial layer flocks. [ABSTRACT FROM AUTHOR]

The exposure assessment study was conducted in the southern Punjab, the cotton growing area of Pakistan among two female groups, one as cotton pickers (occupationally exposed) and the other group as non-pickers. The average blood cholinesterase activity before the spraying in non-pickers were found to be 5.32 ± 0.70 kU/L with the average activity of female cotton pickers 5.31 ± 1.12 kU/L. Whereas, due to environmental contamination the average activity of enzyme cholinesterase (ChE) was calculated to be 4.80 ± 1.08 kU/L in non picker's blood samples, collected after exposure to pesticides (post-season) as compared to 2.81 ± 1.32 kU/L in female cotton pickers. All these values were found be in the normal range. The mean values of ChE were found significantly different (1.95) among various groups when statistically analyzed by Z-test (α = 0.05). The effects of pesticides found equal in both groups of female were discussed in detail with common symptoms observed in majority of cotton pickers. [ABSTRACT FROM AUTHOR]

The mycotoxin ochratoxin A (OTA), a well-known human nephrotoxic and carcinogenic agent, is a public health concern in many countries. Exposure is assessed by means of mycotoxin analysis in food commodities and by human biomonitoring of OTA in blood samples. Data available from several European countries and some studies in Africa, Asia, and the Americas indicate frequent detection of OTA. Thus far, data from developing countries that compare blood levels in healthy and diseased individuals are scarce. Thus, the aim of this investigation was to determine OTA levels in blood samples of bladder cancer patients (n = 96) and healthy controls (n = 31) from Pakistan. OTA in blood plasma was analyzed after extraction by high-performance liquid chromatography (HPLC) with fluorescence detection. Among samples of 87 cancer patients and 30 controls, 92% in total contained quantifiable amounts of OTA. In bladder cancer cases the median OTA concentration was 0.19 ng/ml (mean 0.296; range: 0.03 to 3.41 ng/ml), and in healthy controls the median OTA was 0.19 ng/ml (mean 0.3; range: 0.04 to 1.24 ng/ml). The OTA levels found in the Pakistanian cohorts were comparable to those reported previously for the general population in the European Union. In conclusion, OTA is not likely to play a major role in the etiology of bladder cancer in the Karachi cohort, at least as the sole risk factor. [ABSTRACT FROM PUBLISHER]

Background: To study the role of single nucleotide variants (SNVs) of genes related to preeclampsia in Pakistani pregnant women. Methods: After ethical approval and getting informed consent; 250 pregnant women were enrolled and equally divided into two groups (125 preeclamptic cases and 125 normotensive pregnant women). Demographic details and medical history were recorded, and 10 ml blood sample was obtained for DNA extraction. The tetra-primer amplification refractory mutation system (ARMS) assays were developed for assessing the variants of three preeclampsia related genes; F5, MTHFR and VEGFA. An association of six SNVs; F5:c.1601G > A (rs6025), F5:c.6665A > G (rs6027), MTHFR: c.665C > T (rs1801133), MTHFR: c.1286A > C (rs1801131), VEGFA: c.-2055A > C (rs699947) and VEGFA: c.*237C > T (rs3025039) with preeclampsia was determined by using different genetic models. Results: Genotyping of the SNVs revealed that patients with MTHFR:c.665C > T, have increased susceptibility to preeclampsia (CT versus CC/TT: OR = 2.79, 95% CI = 1.18–6.59; P* = 0.046 and CT/TT vs CC: OR = 2.91, 95% CI = 1.29–6.57; P* = 0.0497, in overdominant and dominant models, respectively), whereas F5:c.6665A > G, (A/G vs AA/GG: OR = 0.42, 95% CI = 0.21–0.84; P* = 0.038 in overdominant model) and MTHFR:c.1286A > C, (CC versus AA: OR = 0.36, 95% CI = 0.18–0.72; P* = 0.0392 in codominant model) have significantly decreased risk for preeclampsia. F5:c.1601G > A, VEGFA: c.-2055A > C and VEGFA: c.*237C > T variants revealed no relationship with the disease. Conclusion: This is the first case control study describing the protective role of F5:c.6665A > G against preeclampsia in any world population. In addition, the present study confirmed the association and role of MTHFR gene variations in the development of preeclampsia in Pakistani patients. Further genetic studies may be required to better understand the complex genetic mechanism of SNVs in preeclampsia related genes in pregnant women. [ABSTRACT FROM AUTHOR]