Molecular prevalence, phylogeny and hematological impact of Toxoplasma gondii and Plasmodium spp. in common quails from Punjab, Pakistan.

This study investigates the molecular prevalence and phylogenetic characteristics of two prominent blood-borne pathogens, Toxoplasma gondii (T. gondii) and Plasmodium spp., in common quails (Coturnix coturnix) sampled from both wild (N = 236) and farmed (N = 197) populations across four districts (Layyah, Dera Ghazi Khan, Lahore, and Multan) in Punjab, Pakistan, during the hunting seasons from 2021 to 2023. Additionally, the impact of these pathogens on the complete blood count (CBC) of the hosts is examined. Out of 433 quails tested, 25 (5.8%) exhibited amplification of the internal transcribed spacer (ITS-1) gene for T. gondii, while 15 (3.5%) showed amplification of the Cytochrome b gene for Plasmodium spp. A risk factor analysis indicated that the prevalence of both pathogens was not confined to specific sampling sites or bird sexes (P > 0.05). District-wise analysis highlighted that hens were more susceptible to both T. gondii and Plasmodium spp. infections than cocks. Wild quails exhibited a higher susceptibility to T. gondii compared to farmed birds. Significant CBC variations were recorded in infected birds as compared to uninfected ones. BLAST analysis of generated sequences has confirmed the identity of recovered PCR amplicons as T. gondii and Plasmodium relictum. Phylogenetic analysis revealed that Pakistani isolates clustered with those reported from various countries globally. This study provides the first documentation of T. gondii and Plasmodium sp. infections in Pakistani quails, underscoring the need for detailed investigations across different regions to enhance our understanding of infection rates and the zoonotic potential of these parasites.

Detection and Phylogenetic Analysis of Extended-Spectrum ß-Lactamase (ESBL)-Genetic Determinants in Gram-Negative Fecal-Microbiota of Wild Birds and Chicken Originated at Trimmu Barrage.

Extended-spectrum ß-lactamases (ESBL) give rise to resistance against penicillin and cephalosporin antibiotics in multiple bacterial species. The present study was conducted to map genetic determinants and related attributes of ESBL-producing bacteria in three wild aquatic bird species and chickens at the "Trimmu Barrage" in district Jhang, Punjab province, Pakistan. To study the prevalence of ESBL-producing bacteria, a total of 280 representative samples were collected from wild bird species; cattle egrets (Bubulcus ibis), little egrets (Egretta garzetta) and common teals (Anas crecca) as well as from indigenous chickens (Gallus gallus domesticus) originating from a local wet market. The isolates were confirmed as ESBL producers using a double disc synergy test (DDST) and bacterial species were identified using API-20E and 20NE strips. A polymerase chain reaction (PCR) was used to detect ESBL genetic determinants and for genus identification via 16S rRNA gene amplification. A phenotypic antimicrobial susceptibility test was performed for ESBL-producing isolates against 12 clinically relevant antibiotics using the Kirby-Bauer disk diffusion susceptibility test. A phylogenetic tree was constructed for the sequence data obtained in this study and comparative sequence data obtained from GenBank. The overall prevalence of ESBL-producing bacteria was 34.64% (97/280). The highest percentage (44.28%; 31/70) of ESBL-producing bacteria was recovered from chickens (Gallus gallus domesticus), followed by little egrets (Egretta garzetta) (41.43%; 29/70), common teal (Anas crecca) (28.57%; 20/70) and cattle egrets (Bubulcus ibis) (24.28%; 17/70). Five different ESBL-producing bacteria were identified biochemically and confirmed via 16S rRNA gene sequencing, which included Escherichia coli (72; 74.23%), Enterobacter cloacae (11; 11.34%), Klebsiella pneumoniae (8; 8.25%), Salmonella enterica (4; 4.12%) and Pseudomonas aeruginosa (2; 2.06%). Based on PCR, the frequency of obtained ESBL genes in 97 isolates was blaCTX-M (51.55%), blaTEM (20.62%), blaOXA (6.18%) and blaSHV (2.06%). In addition, gene combinations blaCTX-M + blaTEM, blaTEM + blaOXA and blaCTX-M + blaSHV were also detected in 16.49%, 2.06% and 1.03% of isolates, respectively. The ESBL gene variation was significant (p = 0.02) in different bacterial species while non-significant in relation to different bird species (p = 0.85). Phylogenetic analysis of amino acid sequence data confirmed the existence of CTX-M-15 and TEM betalactamases. The average susceptibility of the antibiotics panel used was lowest for both Klebsiella pneumoniae (62.5% ± 24.42) and Salmonella enterica (62.5% ± 31.08) as compared to Enterobacter cloacae (65.90% ± 21.62), Pseudomonas aeruginosa (70.83% ± 33.42) and Escherichia coli (73.83% ± 26.19). This study provides insight into the role of aquatic wild birds as reservoirs of ESBL-producing bacteria at Trimmu Barrage, Punjab, Pakistan. Hence, active bio-surveillance and environment preservation actions are necessitated to curb antimicrobial resistance.

Diversity and Abundance of Delineated Earthworm (Annelida: Clitellata) in Pakistan: A Review.

Earthworm, a ubiquitous (but neglected) macro-invertebrate, is found in terrestrial vicinity of Pakistan. Moreover, the occurrence of earthworms is often diverse with fluctuating quantity depending upon abiotic factors and land usage patterns. The aim of this study was to summarize all the reported information related to earthworm diversity in different areas of Pakistan. Almost all the data published from year 2001 to 2021 were collected. Following data organization, total 42 earthworm's species including five families (Acanthodrilidae, Lumbricidae, Moniligastridae, Octochaetidae and Megascolecidae) were reported from various researchers. Among five families, family Acanthodrilidae was found to have only one specie (Ramiella bishambari), Lumbricidae consist of 10 species (Apporactodea rosea, Allolobophora trapezoids, Allolobophora chloroticaa, Aporrectodea longa, A. caliginosa, Bimastus parvus, Eisenia fetida, Helodrilus foetidus, Lumbricus terrestris and L. rubillus), Moniligastridae has two species (Drawida nepalensis and D. pellucida) while Octochaetidae possess only one specie (Eutyphoeus incommodus). The most abundant and diverse family Megascolecidae consist of 28 earthworm species in all habitats of different regions of Pakistan. Among geographical areas, Faisalabad was found as the richest territory with most reported earthworm species (i.e. 28). The current study suggests further in depth research to explore the unidentified and/ missing species of earthworms in Pakistan.

Epidemiology, Diagnosis, and Control of Canine Infectious Cyclic Thrombocytopenia and Granulocytic Anaplasmosis: Emerging Diseases of Veterinary and Public Health Significance.

This review highlights the diagnostic methods used, the control strategies adopted, and the global epidemiological status of canine cyclic thrombocytopenia and granulocytic anaplasmosis at the animal-human interface. Canine anaplasmosis is an important worldwide disease, mainly caused by Anaplasma platys and A. phagocytophilum with zoonotic implications. A. platys chiefly infects platelets in canids, while A. phagocytophilum is the most common zoonotic pathogen infecting neutrophils of various vertebrate hosts. Diagnosis is based on the identification of clinical signs, the recognition of intracellular inclusions observed by microscopic observation of stained blood smear, and/or methods detecting antibodies or nucleic acids, although DNA sequencing is usually required to confirm the pathogenic strain. Serological cross-reactivity is the main problem in serodiagnosis. Prevalence varies from area to area depending on tick exposure. Tetracyclines are significant drugs for human and animal anaplasmosis. No universal vaccine is yet available that protects against diverse geographic strains. The control of canine anaplasmosis therefore relies on the detection of vectors/reservoirs, control of tick vectors, and prevention of iatrogenic/mechanical transmission. The control strategies for human anaplasmosis include reducing high-risk tick contact activities (such as gardening and hiking), careful blood transfusion, by passing immunosuppression, recognizing, and control of reservoirs/vectors.

Appraisal of iron accumulation in soil, forages, and blood plasma of sheep and goats: a case study in different districts of Punjab, Pakistan.

Minerals are essential for ruminants affecting significantly the production of grazing livestock. Iron level in forages, soil, and blood plasma of the small ruminants (goat and sheep) was investigated in three districts of Punjab. Atomic absorption spectrophotometer was used to determine the concentration of iron in collected samples. The results revealed that the mean Fe concentrations in soil of districts Sargodha, Mianwali, and Bhakhar were significantly varied and ranged from 21.85 to 23.78, 28.45 to 31.2, and 18.079 to 24.33 mg/kg, respectively. The Fe level in soil of Mianwali significantly varied and was higher than Sargodha and Bhakkar. The mean Fe concentration in forages which were used for feeding purpose were significantly varied and found between 10.95-14.49, 23.63-25.65, and 6.616-9.45 mg/kg for Sargodha, Mianwali, and Bhakhar, respectively. The mean Fe concentrations in blood plasma of goat which consumed the contaminated forages were 8.5026-11.763 mg/L in district Sargodha, 19.77-20.19 mg/L in Mianwali, and 5.508-5.858mg/L in Bhakkar. In blood plasma of sheep, the residual levels of Fe in districts Sargodha, Mianwali, and Bhakhar were ranged from 9.987 to 12.455, 15.8 to 19.785, and 3.425 to 6.383 mg/L, respectively. This study provides the data of metals effected by different sites and also their mobility from low to higher trophic level which enables us to study the iron toxicity in different trophic levels, and we recommend different safe limits and treatment in case of low and high metal profile.

Hepcidin: A key regulator of iron.

Hapcidin is the central regulator of iron homeostasis in the body. Primarily it is extracted from urine. Hepcidin is a 25 amino acid long chain peptide. Inflammation or iron overload greatly stimulate production of hepcidin by hepatocytes. Recent evidences have revealed that mutations in the human haemochromatosis (HFE) gene lead to deficiency of hepcidin which is responsible for iron overload and contributing to haemochromatosis. Moreover, hepcidin plays a key role in different types of anaemia, mainly anaemia of inflammation in which concentration of hepcidin increases up to 100 folds. Its contribution to renal disease, heart diseases, cancer and obesity-related disorders are also observed. On the other hand, its role is quite inevitable in understanding metastasis in certain cancers. By understanding the mechanism of hepcidin and its pathological roles in blood and iron diseases could lead to new therapies.

Parasitological, serological and molecular survey of Trypanosoma evansi infection in dromedary camels from Cholistan Desert, Pakistan.

BACKGROUND: Surra, a vector borne disease caused by Trypanosoma (T.) evansi, affects the health, productivity and working capacity of camels. Since clinical signs are not pathognomonic, diagnosis must be confirmed by laboratory methods. This is a first study on the prevalence of surra in Cholistan Desert, Pakistan using a broad variety of diagnostic tests thereby emphasizing it as a risk for the dromedaries of Pakistan. METHODS: In a cross sectional study, 1005 dromedary camels from three districts in the Cholistan Desert were sampled to assess the prevalence of trypanosomosis due to T. evansi by means of parasitological (Giemsa stained thin smear), serological (formol gel test, CATT/T. evansi, ELISA/VSG RoTat 1.2, immune trypanolysis) and molecular tests (TBR1/2 PCR and RoTat 1.2 PCR). Kappa was calculated to assess the degree of agreement between different tests whereas chi-square test along with odds ratios and their 95% confidence intervals were used to study influence of breed, gender, age and locality on disease prevalence. RESULTS: Overall prevalence was 0.7% with Giemsa stained thin smears (GST), 40.1% with formol gel test (FGT), 47.7% with CATT/T. evansi, 44.2% with ELISA/VSG RoTat 1.2, 39.9 % with immune trypanolysis (TL), 31.9 % with TBR1/2 PCR and 30.5% with RoTat1.2 PCR. Based on these results, the Cholistan Desert appears to be a high risk area for surra. According to TL and TBR1/2 PCR, camels at Bahawalpur are approximately two times more likely to be infected than those in Bahawalnagar (OR = 1.8; 95% CI: 1.38-2.42) and Rahim Yar Khan (OR = 1.9; 95% CI: 1.30-2.75). Test agreement of TL was moderate with CATT/T. evansi (k = 0.43; 95% CI: 0.378-0.489) and ELISA/VSG RoTat 1.2 (k = 0.54; 95% CI: 0.489-0.594) and poor with the other tests. Test agreement between TBR1/2 PCR and RoTat1.2 PCR was almost perfect (k = 0.96; 95% CI: 0.950-0.984). We didn't find evidence for the presence of T. evansi type B in the studied population. CONCLUSION: Our study supports using antibody detection tests, rather than parasitological and molecular examination, to assess surra prevalence in camels. It also calls for implementation of measures to control surra in the Cholistan Desert.

Histopathological studies on stunting syndrome in broilers, lahore, pakistan.

Runting stunting syndrome (RSS) is a multifactorial disease with many names and faces that had caused considerable economic losses to poultry through reduced uniformity, reduced livability, decreased body weights, elevated feed conversions, and many secondary diseases. The aim of the current study was to evaluate the effect of stunting syndrome on histopathology in chicks (n = 120) of different ages collected from nine different farms. Grouping was done on the basis of age (G1 = 1-10 days, G2 = 11-20 days, G3 = 21-30 days, and G4 = 31-40 days) including both stunted and normal chicks. Histopathological findings were the intestinal lesions (29%), including degeneration of villi, crypts, epithelial cells and lamina propria. Pancreatic histopathological lesions (16.65%) included the fibrosis, vacuolation, and degeneration of acinar cells. Degeneration of follicles and epithelial cells, of bursa of fabricius (43%) and dilation of glandular cells of proventriculus including lymphocytes infiltration (5.6%) were other histopathological findings. All these changes may interfere with normal digestive processes and normal body functioning resulting in poor weight gain and retarded growth or stunting of chicks.