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Recently, there have been numerous reports showing that phthalates have negative human health impacts and may cause several diseases such as asthma, breast cancer, obesity, type II diabetes, and male infertility. Animals are also exposed to phthalates through the environment and can cause adverse health effects on them. Several studies have been found on the cytogenetic effects of dibutyl phthalate (DBP) on different organisms but no documented evidence has been found on the cytotoxic and genotoxic effects of dibutyl phthalate (DBP) on bovine cultured lymphocytes. MTT assay was performed on different series of DBP concentrations (10 μM, 20 μM, 30 μM, 50 μM, 70 μM, 100 μM). A concentration-dependent decrease in cell viability was observed by the DBP. The LD50, LD50/2, and 2∗LD50 were found to be 50 μM, 30 μM, and 80 μM on bovine lymphocytes, respectively. Then, these concentrations of DBP were utilized to perform comet, micronucleus assays, and oxidative stress. A concentration-dependent increase in DNA damage, oxidative stress, and micronuclei formation was observed in lymphocytes by the DBP as compared to the control group. Highest genotoxic effects were observed at a concentration of 2∗LD50. Similarly, total oxidative stress was found higher, and antioxidative stress was lower in concentration-dependent manner by the DBP. The current study revealed a significant cytotoxic, genotoxic, and oxidative stress of DBP on cultured bovine lymphocytes.

The current study evaluated the efficiency of locally characterized recombinant thermostable xylanase (XYLTN) from Thermotoga naphthophila in broiler chicks. The XYLTN was produced using BL21 Codon Plus (DE3) cells having pET-21a containing xylanase gene from Thermotoga naphthophila and was used for the supplementation of poultry feed. For the poultry trail, a total of 150 day old broiler chicks were divided into five groups having 30 birds each. Group A served as negative control while groups B, C, and D were experimental groups and fed on a basal diet supplemented with 1000, 1500, and 2000 IU/Kg of locally produced XYLTN, respectively, whereas group E served as positive control and was fed on diet supplemented with 1500 IU/Kg of commercially available xylanase. The supplementation of poultry feed with XYLTN revealed, a maximum weight gain of 1681.25g, feed intake of 2810g, and feed conversion ratio of 1.67 when the feed was supplemented with 2000 IU/Kg of XYLTN. Locally produced XYLTN exhibited promising outcomes compared to positive control which is being utilized currently for the supplementation of poultry feed in the industry. The weight gain, feed intake, and feed conversion ratio of 1681.25g, 2810g, and 1.67 for experimental group D were comparable to 1610.38g, 2830g, and 1.75 for positive control. The ability of enzyme to enhance weight gain, feed consumption, and feed conversion ratio in poultry chicks makes it a strong candidate for replacement of its commercial counterpart being imported for the poultry industry, and its domestic production will contribute to the economic availability of this xylanase for the poultry feed industry.

ABSTRACT In this study total of 150 Pakistani quail (Coturnix japonica PK) locally evolved in Pakistan were used at the age of 30 days to reveal the relationship between body weight and linear body measurements. Body measurements included body weight (BW), body length (BL), wing spread (WS), shank length (SL), shank circumference (SC), drumstick length (DL), drumstick circumference (DC), breast width (BD) and keel length (KL). The overall association between BW and other body measurements was found highly significant (p-value = 0.000). A multiple linear regression model for both male and female birds was found to be highly significant (p-value=0.000). In male birds, there was a strong positive correlation between BW and BL (p-value=0.000) and a moderate negative correlation between BW and DC (p-value=0.000). Other body measurements were observed as weakly correlated with BW (p values >0.05). In female birds, there was a strong positive correlation between BW and BL (p-value=0.000). The interdependence between BW and SL has been observed to be a moderate negative correlation (p-value < 0.01). The body measurement DC was moderately negatively correlated with body weight BW (p-value < 0.01). Two variables SC and DL were also found to be moderately positively correlated with BW (p values < 0.001 and < 0.004 respectively). The rest of the variables were weakly correlated with BW. This study revealed a strong correlation between BW and BL of both male and female birds which can be further used as criteria for assessment and early selection of Coturnix japonica PK for early body weight evaluation.

Pakistan is bestowed with variety of habitats and climatic conditions which leads to diverse range of avian species. Among avifauna of Pakistan, Birds of prey or raptors are well-known because of their beauty and speed of flight. Falcons, hawks, kites, eagles, and vulture are common birds of prey. They are geographically widespread and common among the vertebrates. Being predatory birds, they are found on the top of food chain. Unfortunately, birds of prey are facing serious threats such as loss of habitat, pollution, poaching and injuries. In order to maintain ecological balance and food chains, this is very important to make strategies for conservation of these predatory birds. However, there is still uncertainty in their taxonomy because these birds are not studied well at genetic level. Morphological identification includes size, color, and body weight etc. which are crude and does not lead to accurate identification at species level. In order to overcome such gaps, the aim of this study was the identification of two broad families of raptors; Accipitridae and Falconidae, at molecular level using mitochondrial ND2 gene. The partial sequence of ND2 gene was submitted to GenBank. The novel SNPs were investigated which serves as marker for identification of Pakistani raptorial species. Two sub species of falcons are also characterized at genetic level for the first time. The study represented the first report on genetic data of raptorial species of the Pakistan. This strategy can be used to identify other species of birds of prey to get diverse genetic data which will be helpful for the conservation planning of these birds. Developed genetic markers of identification will be used for forensic purposes and also play a significant role in maintenance of ecosystems.

ABSTRACT The aim of this study was to develop a criteria on the basis of impact of dengue infection on various clinical, hematological and biochemical parameters in dengue patients for the quick diagnosis and detection of dengue stage for the proper treatment of infection. This cross-sectional study was conducted at Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore and Jinnah Hospital Lahore (JHL) from January to December 2013. Total 345 clinically suspected patients reporting to JHL were serologically diagnosed for dengue specific antigen and antibodies. In this study 20 patients of febrile illness other than Dengue (OFI), were selected as control. The frequency of various clinical features and alterations in hematological and biochemical parameters were assessed for both cases and controls. Total 108 (31.3%) patients were serologically confirmed for dengue infection. Seropositive 108 cases were classified as classical DF 81(75%), DHF 22(20.4%) and DSS 5 (4.6%). Mean age was 32.3±12.4 years, which comprises of male 80 (74%) and female 28 (25.9%). Common symptoms for dengue were fever and headache (100%), arthralgia (82%), myalgia (80.5%), retro-orbital pain (68.5%), bleeding tendencies (38%), rash (51%) and vomiting (48%). Thrombocytopenia (90%), leukopenia (62.5%), elevated transaminases (ALT 56.5% and AST 70.5%), hyponatremia (51.8%), hypokalemia (40.7%) and hypocalcemia (81.4%) was recorded among dengue patient. It was concluded that bleeding tendencies, retro-orbital pain, rash and vomiting were more frequent in DHF and DSS cases. Thrombocytopenia, leukopenia, deranged hematocrit, raised transaminases, urea and creatinine levels and decreased serum levels of albumin, cholesterol, sodium, potassium and calcium were commonly associated with severe form of disease.

Cancer is unregulated growth of cells that can spread to other part of body via blood circulation and lymphatic system. Mutations in genetic material can alter cell physiology, ultimately resulting in tumor. Like human cancers, dogs have relatively high incidence of cancers, relatively large body size and responses to cytotoxic and other therapeutics. Small noncoding RNA having length of 22 base pair (bp) are called micro-RNA (miRNA), that are processed by Dicer from precursors with a characteristic hairpin secondary structure. miRNAs can regulate gene expression at the post-transcriptional level by binding to the 3' untranslated region (UTR) of target mRNAs. In this study dog mammary tumor samples were collected to investigate the expression level of miRNAs hsa-222-3p, hsa-let-7b-5p and hsa-let-7f-5p and effect of their expression on the target genes (HMGA1 and CDKN1B) of these miRNAs. Qiagen miScript Primer Assay based expression analyses revealed the over-expression of all three miRNAs in most of the studied canine mammary tumor samples compared to normal mammary tissue. Furthermore, this over-expression of tested miRNAs, down regulated their target genes in tumor samples compared to normal samples.

Genetic markers, such as single nucleotide polymorphisms (SNPs) in growth hormone (GH) genes, can play a significant role in selection for improving growth traits in poultry birds. The data on SNPs of GH genes of C. japonica in Pakistan is limited. Therefore, this study was conducted to look into the SNPs in GH gene exon 3, 4 and partial sequence of intron 3, 4, and 5 of Pakistani quail Coturnix japonica PK (C. japonica PK) through polymerase chain reaction (PCR). Five body weight categories (upper outliers body weight; > 250g, higher body weight; 211 to 250g, medium body weight: 171 to 210g, small body weight: 130 to 170g and lower outliers body weight; <130g) of birds were made, and GH gene was amplified, sequenced, and screened for detection of SNPs. Furthermore, polymorphisms and their association with different growth traits such as body weight (BW), body length (BL), wing spread (WS), shank length (SL), shank circumference (SC), drumstick length (DL), drumstick circumference (DC), breast width (BD) and keel length (KL) of C. japonica PK were also evaluated. A total of eighteen SNPs were detected in the GH gene. Genotypes of three SNPs were significantly associated with BW. Genotype TC at position c.31T>C (exon 3) and genotype AA at position c.161A>C (exon 3) had greater BW as compared to the other genotypes (P=0.000). Whereas n.703G>A (intron 4) revealed a very strong association of genotype (AA) with breast width (BD) (P=0.001) and n.12C>T (intron 5) revealed a very strong association of genotypes (CC and TT) with large body weight category and upper outlier body weight category (P=0.008) and DC (P=0.038). c.31T>C (exon 3) revealed a very strong association of genotype (TC) with upper outlier body weight categories (P=0.000), BL (P=0.000) and, DL (P=0.004). c.161A>C (exon 3) revealed a very strong association of genotype (AA) with upper outlier body weight categories (P=0.000), SC (P=0.040) DC (P=0.032) and BW (P=0.007). Thus, GH gene may be used as a candidate gene, to improve growth traits of C. japonica PK. These results demonstrated that these three SNPs of the GH gene might be exploited further as a candidate genetic marker for the genetic improvement of growth-related traits in C. japonica PK.

ABSTRACT Salmonella enterica subsp. enterica poses a threat to both human and animal health, with more than 2500 reported serovars. A total of 80 samples, comprising of poultry meat (n=30) from poultry shops and supermarkets, poultry feed (n=30) and swabs from carcasses and muddy (n=20) of poultry shops. The samples were assessed microbiologically for Total Viable Count, Total Coliform Count, and Salmonella detection. The mean log values of total viable counts of meat samples of traditional poultry shops, super markets and processed meat were 5.70, 4.65 and 3.60, respectively and significant (p < 0.05) results were obtained. The mean log values of total coliform counts in meat samples were 2.7, 2.31 and 2.11, respectively. E. coli was predominant 73% in coliform count of all samples. Salmonella was found in 3.75% of samples in which retail poultry shops showed 10%, supermarkets showed 10%. While, processed meat was found negative for Salmonella. The mean log values of total viable counts of feed samples of store and shed were 7.21 and 7.56, respectively. Results of present study showed absence of Salmonella and coliform bacteria in poultry feed samples collected from poultry shed and store-room of poultry farm. Out of 20 swabs only 5% showed Salmonella prevalence. Molecular detection of Salmonella in collected meat samples through PCR targeting His-J gene showed 6.66% of positive samples previously identified by culturing and biochemical profile. The study showed that poultry meat has highest bacterial load which reflects unsatisfactory sanitation and hygienic conditions in poultry environment that ultimately cause food-borne infections. Besides this, feed also becomes a source of bacterial contamination in animals and humans. This study was helpful in devising strategy to provide safe food for public consumption.

ABSTRACT Bovine herpesvirus 1 (BoHV-1) is an emerging disease responsible for high production losses in the dairy animals. A cross-sectional study was conducted to determine the seroprevalence of BoHV-1 in and associated risk factors with disease prevalence in various agro-ecological zones of Pakistan focusing on the Punjab Province. A total of 400 samples were randomly collected from cattle and buffalo of wheat rice Punjab, mixed Punjab, cotton/wheat Punjab, Low intensity Punjab and Rain-fed Punjab zones. The study revealed individual seroprevalence of BoHV-1 49.5% in cattle and 51.5% in buffalo. Over all prevalence in five zones of Punjab was observed as 25% in mixed Punjab to 71.25% in Rain-fed Punjab. The BoHV-1 seropositivity ranged from 30 - 60% in cattle and 20-75% in buffaloes. Age, breeds of animals and milking status were not found to be potential risk factors for the disease. Logistic regression demonstrated that herd size (>thirty), parity (>2) and abortion history are potential risk factors associated with seroconversion of BoHV-1.

Breast cancer is one of the most common cancers among Pakistani women. It is mostly diagnosed at stage 2, requiring chemotherapy in certain cases. Chemotherapy is of two types: adjuvant and neoadjuvant. It can be recommended at any stage of cancer, either in early stages to shrink the tumor or in late stages to improve quality of life. The side effects may include weight loss, fatigue, hair loss, sores in the mouth, loss of appetite, nausea, vomiting, difficulty swallowing, throat problems, diarrhea, infections, and anemia, which can deteriorate health. Some women may experience weight gain during treatment, which is a risk factor for cancer recurrence. Chemotherapy has many side effects that can directly affect dietary patterns and worsen nutritional status. This study aimed to evaluate the changes in dietary patterns of breast cancer patients during and after chemotherapy and assess how the side effects of chemotherapy affect their nutritional status. Data were collected from 200 breast cancer patients undergoing chemotherapy at Jinnah Hospital, Lahore, using a questionnaire and the Patient-Generated Subjective Global Assessment (PG-SGA) screening tool. The questionnaire included sociodemographic information, questions about disease, treatment, and dietary recall. PG-SGA was used to screen for malnutrition and assess nutritional status. Data collection occurred from September to December 2023. Most participants were female (99%), married (73.5%), and of low socioeconomic status (60.5%). The common stage of cancer was stage 3, and neoadjuvant chemotherapy was recommended for most participants. Effects on body weight varied, with some experiencing weight loss, others weight gain, and some no change. Side effects such as vomiting, nausea, diarrhea, dry mouth, loss of appetite, altered taste and smell, and anemia were commonly reported. Most participants were moderately malnourished or suspected of malnutrition and needed intervention for symptom management. A majority (82%) were overweight or obese according to BMI, indicating a higher risk of cancer recurrence. Participants' dietary intake mainly consisted of carbohydrate- and protein-rich foods during treatment. The side effects of chemotherapy varied among patients. The impact on body weight also differed, with some experiencing weight loss, others weight gain, and some no change. Regarding nutritional status, most patients were moderately malnourished, while a few were severely malnourished.