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Mastitis is one of the major prevailing and costly diseases in dairy cattle and buffaloes. This study describes the current prevalence of mastitis in lactating cattle, and buffaloes in Khyber Pakhtunkhwa during 2019-2020, to identify bacterial pathogens of mastitis, and to determine the antimicrobial resistance. A total of 1013 milk samples belonging to 726 cattle and 287 buffaloes were examined to detect clinical mastitis (CM) and subclinical mastitis (SCM). Mastitic milk samples (n=541) were further used to investigate the prevalence of bacteria associated with mastitis and their antimicrobial susceptibilities by standard disc diffusion assay. The results showed that the prevalence of CM and SCM was 21.2% and 62.3% in cattle, while 17.8% and 68.9% in buffaloes, respectively. Additionally, it was noted that CM was higher in cattle, while SCM was higher in buffaloes. Amongst the bacterial isolates from mastitic milk the prevalence of Staphylococci spp. (24.6%) was at highest, which was followed by Escherichia coli (23.7%), then Streptococci spp. (8%) Salmonella spp. (5.9%), Klebsiella spp. (5.7%), Candida spp. (4.1%), and Proteus spp. (1.5%). The isolated bacteria were mostly resistant to ampicillin (96.5%), sulphamethoxazole (96.5%), streptomycin (95.1%), oxytetracycline (85%), and amoxicillin (78.1%). However, the isolates were highly susceptible to enrofloxacin (86.2%), gentamicin (83.5%), and florfenicol (82.6%). This study might be helpful to the clinicians and researchers associated with the dairy industry for designing prophylactic as well as therapeutic strategies for the control of bovine mastitis. [ABSTRACT FROM AUTHOR]

Simple Summary: Livestock production needs to be improved to ensure global food security and meet the growing demand for food. A sheep's gestation length (the period from conception to birth) is usually about 150 days. We have discovered a novel sheep breed in Chitral, Pakistan, whose individuals may gestate for a relatively shorter period, i.e., 100 days, and yet can give birth to normal lambs. To explore the underlying genetic causes for this unique character we used DNA markers. This approach helped us identify some contrasting genetic variations among short-gestating (Kari-S) and long-gestating (Kari-L) Kari ewes. Two of the total 31 targeted DNA sites demonstrated an association with gestation length in Kari sheep. Following an in-depth analysis and further confirmation in other sheep breeds, these DNA sites may be linked to genes controlling gestating length in sheep. The findings from this study may open ways for faster sheep reproduction with reduced cost. Kari sheep inhabiting the Chitral district of Pakistan show variation in gestation length. In this study, we have analyzed the genetic differences between the three subtypes of Kari sheep (based on variation in gestation length) using microsatellite markers. Kari sheep samples were collected from their breeding tract and were characterized for gestation length and genetic diversity using microsatellite markers. A total of 78 Kari ewes were grouped into three categories based on gestation length (GL), i.e., Kari-S (with a shorter GL), Kari-M (with a medium GL), and Kari-L (with a longer GL). DNA from these samples was used to amplify 31 ovine-specific microsatellite loci through PCR. Of the total 78 Kari specimens, 24 were grouped in Kari-S (GL = 100.7 ± 1.8), 26 were from the Kari-M subtype (GL = 123.1 ± 1.0), and 28 were Kari-L (GL = 143.8 ± 1.5). Microsatellite analysis revealed an association of genotypes at two marker sites (MAF214 and ILSTS5) with variation in GL. A total of 158 alleles were detected across the 22 polymorphic loci with an average of 7.18 alleles per locus. Unique alleles were found in all three subtypes. The highest number of unique alleles was observed in Kari-L (15), followed by Kari-S (10) and Kari-M (8). The results indicated that Kari-S is a genetically distinct subtype (with higher genetic differentiation and distance) from Kari-M and Kari-L. The genetic uniqueness of Kari-S is important for further exploration of the genetic basis for shorter gestation length, and exploitation of their unique values. [ABSTRACT FROM AUTHOR]

Objective: To determine the association between epilepsy disease (ED) and consanguinity in the tribal society of Khyber Pakhtunkhwa-Pakistan. Material and methods: Approximately, 150 clinically diagnosed epileptic patients of consanguineous families with ages ranging from 3 to 70 years were included during the period of (2018-19). Children in this study were aged 7-10 years and male to female ratio among the patients was 1:1.5. The target familial cases were included on the basis of sex. age. parental consanguinity, seizure type, cause of epilepsy, and family history. The parents were interviewed to collect the required information after informed consent. Non-consanguineous families, patients with acquired causes, and those who were not willing to participate were excluded from the study. Results: In the current study, the consanguinity in parents of the epileptic patients was compared with the general population and we found significant results (P< 0.01). Generally, 56 (37.33%) of the patient's parents were first cousins with (OR=3.245 and 95%-CI =2.26-4.645 and P<0.0001), 24(16.00%) were second-degree cousins with (OR=3.269 and 95%-CI=2.032- 5.258 and P<0.0001) and 70 (46.66%) were unrelated. Conclusion: The overall results in this study clearly indicate the potential role of heritage lineage/consanguinity in epilepsy within consanguineous families. The present retrospective study supports genetic counselling prior to marriages. More significant genotype-phenotype correlation is also needed to minimize the risk of epilepsy disease. [ABSTRACT FROM AUTHOR]

Prion infections of the central nervous system (CNS) are characterized by initial reactive gliosis followed by overt neuronal death. Gliosis is likely to be caused initially by the deposition of misfolded, proteinase K-resistant, isoforms (termed PrPSc) of the normal cellular prion protein (PrPc) in the brain. Proinflammatory cytokines and chemokines released by PrPSc-activated glia and stressed neurons may also contribute directly or indirectly to the disease development by enhancing gliosis and inducing neurotoxicity. Recent studies have illustrated that early neuroinflammation activates nuclear factor of activated T cells (NFAT) in the calcineurin signaling cascade, resulting in nuclear translocation of nuclear factor kappa B (NF-κB) to promote apoptosis. Hence, useful therapeutic approaches to slow down the course of prion disease development should control early inflammatory responses to suppress NFAT signaling. Here we used a hamster model of prion diseases to test, for the first time, the neuroprotective and NFAT-suppressive effect of a second-generation semisynthetic tetracycline derivative, minocycline, versus a calcineurin inhibitor, FK506, with known NFAT suppressive activity. Our results indicate that prolonged treatment with minocycline, starting from the presymptomatic stage of prion disease was more effective than FK506 given either during the presymptomatic or symptomatic stage of prion disease. Specifically, minocycline treatment reduced the expression of the astrocyte activation marker glial fibrillary acidic protein and of the microglial activation marker ionized calcium-binding adapter molecule-1, subsequently reducing the level of proinflammatory cytokines interleukin 1β and tumor necrosis factor-α. We further found that minocycline and FK506 treatment inhibited mitogen-activated protein kinase p38 phosphorylation and NF-κB nuclear translocation in a caspase-dependent manner, and enhanced phosphorylated cyclic adenosine monophosphate response element-binding protein and phosphorylated Bcl2-associated death promoter levels to reduce cognitive impairment and apoptosis. Taken together, our results indicate that minocycline is a better choice for prolonged use in prion diseases and encourage its further clinical development as a possible treatment for this disease. [ABSTRACT FROM AUTHOR]

Injection drug users (IDUs) are considered as a high risk group to develop hepatitis C due to needle sharing. In this study we have examined 200 injection drug users from various regions of the Khyber Pakhtunkhwa province for the prevalence of active HCV infection and HCV genotypes by Immunochromatographic assays, RT-PCR and Typespecific PCR. Our results indicated that 24% of the IDUs were actively infected with HCV while anti HCV was detected among 31.5% cases. Prevalent HCV genotypes were HCV 2a, 3a, 4 and 1a. Majority of the IDUs were married and had attained primary or middle school education. 95% of the IDUs had a previous history of needle sharing. Our study indicates that the rate of active HCV infection among the IDUs is higher with comparatively more prevalence of the rarely found HCV types in KPK. The predominant mode of HCV transmission turned out to be needle sharing among the IDUs. [ABSTRACT FROM AUTHOR]

Estrogen biosynthesis is catalyzed by the Aromatase (P450arom), which is derived from CYP19A1. We investigated the relationship between actin filament re- arrangement, CYP19A1 expression and estradiol bio-synthesis in bovine granulosa cells in vitro. Both bovine fibroblasts and granulosa cells were starved in serum-free culture medium for 24 hours. Serum-starvation disassembled stress fibers in fibroblasts, but not in granulosa cells. Additionally we added Y27632 and CB to medium to de-polymerize the actin filaments, while LPA and insulin were used to re-polymerize actin filaments. A real-time RT-PCR analysis revealed that, FSH induced transcription of CYP19A1 mRNA encoded aromatase was inhibited significantly as actin filaments were de-polymerized by both Y27632 and CB, whereas, restored as actin filaments were re-polymerized by both LPA and insulin. However, further study shows that, statistically there were no differences in the expression of aromatase proteins analyzed by western-blot, during the process of actin filament rearrangement. ARIA analysis also indicated that the secretions of estradiol by bovine granulosa cells were not changed significantly. The results suggested that serum-starvation could not induce actin filaments depolymerization in bovine granulosa cells, while chemically induced actin filament rearrangement influenced the expression of aromatase in mRNA level. But there were no differences in protein level and biological activity of bovine granulosa cells in vitro. [ABSTRACT FROM AUTHOR]

The present study was designed to establish transgenic Eimeria mitis (tE. mitis) expressing chicken interleukin-4 (ChIL-4) and further exploitation of tE. mitis as a novel vaccine vector to control coccidiosis. For this purpose we compared the clinico-hematological alterations induced by wild type and tE. mitis carrying ChIL- 4 in broilers birds. Wild type E. mitis (wE. mitis) oocysts were nucleofected with double cassette vector and transgenic oocysts were propagated for stable transfection. Stability of the transfected E. mitis was observed by fluorescent microscopy and PCR analysis revealed the appearance of 411bp band suggestive of presence of IL-4 into the genome of tE. mitis. Birds inoculated with transgenic EmiChIL-4 excreted fewer oocysts and fecundity potential of tE. mitis turned out to be significantly lower than that of the wild type parent strain. Moderate clinical and behavioral signs such as restlessness, diarrhea, emaciation, decreased feed intake and loss of body weight was observed in birds inoculated with wE. mitis oocysts. The erythrocyte counts, Hb conc, PCV and MCHC decreased in birds infected with wE. mitis as compared to those infected with tE. mitis. The total leukocyte count including heterophils, eosinophil and basophils increased while lymphocytes were decreased in birds inoculated with wE. mitis as compared to birds in control group and those infected with tE.mitis. It was concluded that tE. mitis incorporated with ChIL-4 functions well as an adjuvant, conferring reduced fertility and compromised pathogenicity of tE. mitis. This discrimination, may be applied as a potential vaccine vector to control coccidiosis as well as pathogens from which antigens are expressed by tE. mitis. [ABSTRACT FROM AUTHOR]

We report the genetic analysis of two consanguineous pedigrees of Pakistani ancestry in which two siblings in each family exhibited developmental delay, epilepsy, intellectual disability and aggressive behavior. Whole-genome sequencing was performed in Family 1, and we identified ~80,000 variants located in regions of homozygosity. Of these, 615 variants had a minor allele frequency ≤ 0.001, and 21 variants had CADD scores ≥ 15. Four homozygous exonic variants were identified in both affected siblings: PDZD7 (c.1348&#95;1350delGAG, p.Glu450del), ALG6 (c.1033G>C, p.Glu345Gln), RBM20 (c.1587C>G, p.Ser529Arg), and CNTNAP2 (c.785G>A, p.Gly228Arg). Sanger sequencing revealed co-segregation of the PDZD7, RBM20 , and CNTNAP2 variants with disease in Family 1. Pathogenic variants in PDZD7 and RBM20 are associated with autosomal recessive non-syndromic hearing loss and autosomal dominant dilated cardiomyopathy, respectively, suggesting that these variants are unlikely likely to contribute to the clinical presentation. Gene panel analysis was performed on the two affected siblings in Family 2, and they were found to also be homozygous for the p.Gly228Arg CNTNAP2 variant. Together these families provide a LOD score 2.9 toward p.Gly228Arg CNTNAP2 being a completely penetrant recessive cause of this disease. The clinical presentation of the affected siblings in both families is also consistent with previous reports from individuals with homozygous CNTNAP2 variants where at least one allele was a nonsense variant, frameshift or small deletion. Our data suggests that homozygous CNTNAP2 missense variants can also contribute to disease, thereby expanding the genetic landscape of CNTNAP2 dysfunction., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Badshah, Mattison, Ahmad, Chopra, Johnston, Ahmad, Khan, Sarwar, Cutler, Taylor, Vadlamani, Zwick and Escayg.)

A captive 8 yr old male bottlenose dolphin Tursiops truncatus succumbed to septicemia with multisystemic inflammation including suppurative enteritis, encephalitis, and pneumonia with chronic pancreatitis. A pure culture of beta-hemolytic, catalase- and oxidase-negative, Gram-positive cocci was isolated from the hilar lymph nodes and pancreas. The isolate was identified by 16S rDNA sequencing as Streptococcus iniae. Histological examination of the digestive system revealed a mixed infection of both bacteria and fungus. Recognized as a pathogen in fish, dolphins, and humans, this is the first report of S. iniae in a dolphin in mainland China. As the number of managed animals in oceanariums is increasing, so is the frequency of contact with fish used as food for marine mammals and humans, highlighting the importance of education and appropriate personal protective protocols to minimize the risk of transmission. An understanding of marine mammal infectious disease organisms is essential to ensuring the health of marine mammals and humans coming into contact with such animals and their food. This study illustrates a systematic clinical, microbiological, and pathological investigation into a septicemic bottlenose dolphin infected with S. iniae. Our findings provide useful information for those involved in the diagnosis and control of infectious diseases in marine mammals and offer insight into an important zoonotic pathogen.

Injection drug users (IDUs) are considered as a high risk group to develop hepatitis C due to needle sharing. In this study we have examined 200 injection drug users from various regions of the Khyber Pakhtunkhwa province for the prevalence of active HCV infection and HCV genotypes by Immunochromatographic assays, RT-PCR and Type-specific PCR. Our results indicated that 24% of the IDUs were actively infected with HCV while anti HCV was detected among 31.5% cases. Prevalent HCV genotypes were HCV 2a, 3a, 4 and 1a. Majority of the IDUs were married and had attained primary or middle school education. 95% of the IDUs had a previous history of needle sharing. Our study indicates that the rate of active HCV infection among the IDUs is higher with comparatively more prevalence of the rarely found HCV types in KPK. The predominant mode of HCV transmission turned out to be needle sharing among the IDUs.