Novel insights regarding the safety and efficacy of pyrethroid-coated nanoparticles against Hyalomma ticks.

Nanoparticles have been shown to inhibit major life cycle stages of ticks, indicative of the promising application of nanomaterials against hard ticks. The study thus probed into one of the alternative options to curtail Hyalomma by employing nanocomposites consisting of pyrethroids (cypermethrin and deltamethrin) coated nanoparticles of iron oxides and iron sulfides keeping alongside the evaluation of their toxicity through plant and mammalian cell lines. The nanoparticles used in this study were roughly spherical in morphology and exhibited various size dimensions upon characterization using SEM, EDX, and FTIR. The application of nanomaterials on female ovipositioning tick showed a decline up to 15% (females ovipositioned) in deltamethrin-coated FeO NPs, whereas this decline was up to 18% in Cyp-FeS NPs and up to 5% in Cyp-FeO NPs. Similarly, the larval hatching was also impacted, leading to a hatching percentage of 5% and only 1% by application of Cyp-FeS NPs and Cyp-FeO NPs, respectively. Similarly, the larval groups had LC90 of 4.1 and 4.73 mg/L for the Cyp-FeO NPs and Cyp-FeS NPs groups. The delta-FeO NPs and delta-FeS NPs demonstrated a promising effect against adult ticks, showing LC50= 3.5 mg/L, LC90= 6.7 mg/L and LC50= 3.8 mg/L, LC90= 7.9 mg/L, respectively. MTT assay revealed that the pyrethroids coupled with iron oxide nanoparticles showed the least cytotoxicity even at the highest concentration (10-1 µL) among other nanomaterials. The study thus concluded a safer spectrum of non-target effects of pyrethroids-coated nanomaterials in addition to their significant anti-tick activity.

Toxocara canis seropositivity in different exposure groups in the Khyber Pakhtunkhwa province of Northwest Pakistan.

Human toxocariasis is a highly prevalent zoonosis worldwide but is underreported in most countries. This study was conducted to evaluate Toxocara canis seropositivity in different exposure sub-groups located in the Mardan, Swabi, and Nowshera districts of the Khyber Pakhtunkhwa province of Northwest Pakistan. A total of 400 blood samples were collected from males 15 years of age and older with no animals, with livestock, with dogs and/or cats living in the house, and from butchers and veterinarians or para-veterinarians. Serum was tested using a commercial ELISA kit for detection of IgG antibodies against T. canis. Proportion seropositive was presented for each group and differences between groups were evaluated using the chi-square or Fisher's exact test as appropriate. Possible risk factors obtained through administration of a questionnaire were also evaluated for each sub-population. Overall T. canis seroprevalence was 14.2%, with a significant difference found between the seroprevalence of individuals with no animals (5.0%; 5/100), individuals with dogs and/or cats living in the household (8.0%; 8/100), individuals with livestock (18.0%; 18/100), veterinarians or para-veterinarians (24.0%; 12/50), and butchers (28.0%; 14/50) (p < 0.001). Significant differences in seropositivity by income bracket, education level, and working in the fields were found for some sub-groups. Study findings demonstrate that certain sub-populations, in Northwest Pakistan, may be at greater risk of T. canis infection. Development and implementation of targeted preventive strategies may, therefore, be needed.

Vaccines against chicken coccidiosis with particular reference to previous decade: progress, challenges, and opportunities.

Chicken coccidiosis is an economically significant disease of commercial chicken industry accounting for losses of more than £10.4 billion (according to 2016 prices). Additionally, the costs incurred in prophylaxis and therapeutics against chicken coccidiosis in developing countries (for instance Pakistan according to 2018 prices) reached US $45,000.00 while production losses for various categories of chicken ranges 104.74 to US $2,750,779.00. The infection has been reported from all types of commercial chickens (broiler, layer, breeder) having a range of reported prevalence of 7-90%. The concern of resistance towards major anticoccidials has provided a way forward to vaccine research and development. For prophylaxis of chicken coccidiosis, live virulent, attenuated, ionophore tolerant strains and recombinant vaccines have been extensively trialed and commercialized. Eimeria antigens and novel vaccine adjuvants have elicited the protective efficacy against coccidial challenge. The cost of production and achieving robust immune responses in birds are major challenges for commercial vaccine production. In the future, research should be focused on the development of multivalent anticoccidial vaccines for commercial poultry. Efforts should also be made on the discovery of novel antigens for incorporation into vaccine designs which might be more effective against multiple Eimeria species. This review presents a recap to the overall progress against chicken Eimeria with particular reference to previous decade. The article presents critical analysis of potential areas for future research in chicken Eimeria vaccine development.

Moringa oleifera alleviated oxidative stress, physiological and molecular disruption induced by acute thermal stress in grass carp, Ctenopharyngodon idella.

The present study was designed to investigate the protective effect of dietary Moringa oleifera leaf meal (MLM) supplementation against high temperature-induced stress in grass carp (Ctenopharyngodon idella). A total of 180 apparent healthy juvenile grass carp (15.48 g ± 0.054) were divided into three groups in triplicate (20 fish in each replicate). Fish were fed with diets containing 0, 1, and 5% MLM for 60 days and then subjected to a high-temperature challenge for 48 h (32-33 °C). The results revealed that feeding fish with 1 and 5% MLM resulted in a significant increase in weight gain and specific growth rate compared to the control. In addition, feed conversion ratio was significantly reduced in groups fed with MLM. No significant difference was reported in the serum cortisol level among different experimental groups before heat stress while serum glucose level significantly decreased in fish fed with 5% MLM. Serum alanine transaminase, aspartate transaminase, and alkaline phosphatase significantly decreased in fish fed with 1 and 5% MLM before and after heat stress. Hepatic lipid peroxidation significantly decreased in fish fed with MLM for 60 days. A non-significant increase in hepatic reduced glutathione level was reported in fish fed with 1 and 5% MLM before heat stress. Catalase and superoxide dismutase activities increased significantly in the liver of fish fed with 5% MLM. No significant change was observed in the expression profile of heat shock protein (hsp) 70 and 90 before heat stress. Meanwhile, after heat stress, up to a fivefold increase was recorded in mRNA level of hsp 70 and fourfold increase in the expression level of hsp 90 in the liver of the control fish which were not fed with MLM-supplemented diets. Fish fed with 1 and 5% MLM showed a significant decrease in the expression of hsp 70 and a non-significant decrease in the expression of hsp 90. Results of the present study suggest that supplementing the diet of grass carp with 5% MLM could improve growth and physiological performance and provide resistance against high temperature-induced stress.

First report on molecular surveillance based on duplex detection of Anaplasma marginale and Theileria annulata in dairy cattle from Punjab, Pakistan.

Theileriosis and anaplasmosis are important tick-borne hemoparasites of bovines. The first surveillance study aimed to assess the suitability of duplex PCR for simultaneous detection of Theileria annulata and Anaplasma marginale field infections in Jhang and Rawalpindi districts of Punjab, Pakistan. Cattle blood samples (n = 480) were collected from selected union councils of all tehsils using a multistage sampling technique. The sampling unit consisted of asymptomatic cattle belonging to either age, sex, and breed. Epidemiological data related to host, area, management, and season were collected using a questionnaire. Based on duplex PCR, the overall prevalence of the two concurrent tick-borne pathogens was 19.79% (95/480). Chi-square analysis indicated that age, breed, tick infestation, history of tick-borne diseases, frequency of acaricidial application, and season were significantly associated with tick-borne pathogens. Phylogenetic analysis of A. marginale and T. annulata isolates based on msp1ß and cytochrome b genes, respectively, revealed that nucleotide sequences acquired from these two pathogens are novel, grouped separately from different countries. All our A. marginale isolates showed 88.2 to 80.5% similarity with isolates from Egypt, Israel, Mexico, and lesser homology with South African isolates. Similarly, the phylogenetic tree based on cytochrome b partial sequences of T. annulata revealed that our sequences are closer to those from India and Iran. Based on this first study on concomitant detection of tick-borne pathogens, it can be concluded that mixed infections are endemic in the study districts and mPCR is suitable for detecting concurrent field infections. Simultaneous infections should be considered while performing surveillance and chemotherapeutic trials for better prevention and control of tick-borne diseases.

Aflatoxins associated oxidative stress and immunological alterations are mitigated by dietary supplementation of Pichia kudriavzevii in broiler chicks.

Mycotoxins are the secondary metabolites of certain toxigenic fungi which pose severe health stress in humans, animals and poultry. Certain biological agents and components are used to adsorb mycotoxins in poultry industry which provide promising results in this regard. Pichia kudriazevii (PK), a novel yeast, has the ability to enhance the immune status of poultry chicks. The present study was designed to investigate the ameliorative potential of PK against aflatoxins associated immunosuppression and oxidative stress in broiler chicks. 180-one day old broiler chicks were equally divided into six groups and given different combinations of aflatoxins (300 and 600 µg/kg) and PK (1 g/kg). Parameters studied were antibody response to sheep red blood cells, lymphoproliferative response to PHA-P; phagocytic response by carbon clearance assay system, total antioxidant capacity and total oxidant status of chicks. Results of this experiment confirmed the immunomodulation and antioxidant capacity of PK against 300 µg/kg aflatoxin level. However such amelioration was partial when PK was used with 600 µg/kg aflatoxins. Moreover, the exact ratio of aflatoxin: PK to cause such amelioration still needs to be investigated.

Helminth protection against type-1 diabetes: an insight into immunomodulatory effect of helminth-induced infection.

Helminths are the old dirty friends of humans from decades and may live undetected by the immune system for years in the tissues. They have evolved as good experts at subverting the immune system. Despite of their pathogenicity, they provide protection to their host against certain inflammatory diseases such as diabetes by modulating the immune mechanisms. These parasites are extra-cellular and induce Th2 response which triggers the adaptive immune cells as well as innate immune cells to work synergistically allowing Tregs to work in a toll-like receptor-dependent manure. T-helper cells type-2 also secrete certain anti-inflammatory cytokines including IL-4, IL-10, IL-13 and TGF-ß which also provide protection against type-1 diabetes. Several helminths such as T. crassiceps, S. venezuelensis, filarial worms, Schistosoma spp. and T. spiralis have been reported to prevent diabetes in mouse models as well as in some clinical trials. Immunomodulatory talent of helminths is receiving greater attention to prevent diabetes. Herein, an attempt has been made to review and highlight the possible immuno-modulatory mechanisms by which helminths provide protection against diabetes. Moreover, this review also emphasizes on the use of helminth-derived molecules or synthetic derivatives of helminth-antigens in clinical trials to overcome rapidly growing autoimmune disorders including diabetes.

Long-term exposure to the fluoride blocks the development of chondrocytes in the ducks: The molecular mechanism of fluoride regulating autophagy and apoptosis.

Long-term exposure to excessive fluoride causes chronic damage in the body tissues and could lead to skeletal and dental fluorosis. Cartilage damage caused by excessive fluoride intake has gained wide attention, but how fluoride accumulation blocks the development of chondrocytes is still unclear. Here, we report a negative correlation between the length and growth plate width after NaF treatments via apoptosis and autophagy, with shrinkage of cells, nuclear retraction, dissolution of chondrocytes. Whereas, fluoride exposure had no significant effect on the number and distribution of the osteoclasts which were well aligned. More importantly, fluoride exposure induced apoptosis of tibial bone through CytC/Bcl-2/P53 pathways via targeting Caspase3, Caspase9, Bak1, and Bax expressions. Meanwhile, the Beclin1, mTOR, Pakin, Pink, and p62 were elevated in NaF treatment group, which indicated that long-term excessive fluoride triggered the autophagy in the tibial bone and produced the chondrocyte injury. Altogether, fluoride exposure induced the chondrocyte injury by regulating the autophagy and apoptosis in the tibial bone of ducks, which demonstrates that fluoride exposure is a risk factor for cartilage development. These findings revealed the essential role of CytC/Bcl-2/P53 pathways in long-term exposure to fluoride pollution and block the development of chondrocytes in ducks, and CytC/Bcl-2/P53 can be targeted to prevent fluoride induced chondrocyte injury.

COVID-19 and Ivermectin: Potential threats associated with human use.

Drugs re-purposing due to COVID-19 virus has declared a number of useful candidates for treatment and prevention of the virus. Ivermectin (IVM) has gained much popularity due to a strong background of magical applications against a broad spectrum of pathogens. The in- vitro studies of ivermectin have shown promise, the thorough clinical trials of its efficacy in the treatment and prevention of SARS-CoV-2 are still warranted. Useful strategies for analyzing projected use of IVM in human coronaviruses might be developed. It may be done by concluding ongoing clinical trials and culturing lessons from IVM usage in veterinary practice. The potential toxicity and careful dosage analyses are urgently required before declaring it as an anti-SARS-CoV-2 drug candidate. This manuscript overviews the background and potential threats associated with the off-label use of IVM as prophylactic drug or treatment option against COVID-19 virus.

Gut microbial dysbiosis and its association with esophageal cancer.

Due to its aggressive nature and low survival rate, esophageal cancer is one of the deadliest cancer. While the intestinal microbiome significantly influences human health and disease. This research aimed to investigate and characterize the relative abundance of intestinal bacterial composition in esophageal cancer patients. The fecal samples were collected from esophageal cancer patients (n = 15) and healthy volunteers (n = 10). The PCR-DGGE was carried out by focusing on the V3 region of the 16S rRNA gene, and qPCR was performed for Bacteroides vulgatus, Escherichia coli, Bifidobacterium, Clostridium leptum and Lactobacillus. High-throughput sequencing of the 16S rRNA gene targeting the V3+V4 region was performed on 20 randomly selected samples. PCR-DGGE and High-throughput diversity results showed a significant alteration of gut bacterial composition between the experimental and control groups, which indicates the gut microbial dysbiosis in esophageal cancer patients. At the phylum level, there was significant enrichment of Bacteroidetes, while a non-significant decrease of Firmicutes in the experimental group. At family statistics, a significantly higher level of Bacteroidaceae and Enterobacteriaceae, while a significantly lower abundance of Prevotellaceae and Veillonellaceae were observed. There was a significantly high prevalence of genera Bacteroides, Escherichia-Shigella, while a significantly lower abundance of Prevotella_9 and Dialister in the experimental group as compared to the control group. Furthermore, the species analysis also showed significantly raised level of Bacteroides vulgatus and Escherichia coli in the experimental group. These findings revealed a significant gut microbial dysbiosis in esophageal cancer patients. So, the current study can be used for the understanding of esophageal cancer treatment, disease pathway, mechanism, and probiotic development.

Pathophysiology of peste des petits ruminants in sheep (Dorper & Kajli) and goats (Boer & Beetal).

Peste des petits ruminants (PPR), an economically important viral transboundary disease of small ruminants is not only prevalent in Pakistan but also in other countries where people rely on agriculture and animal products. The present study was aimed at describing the pathology and antigen localization in natural PPR infections in local (Kajli sheep; Beetal goats) as well as imported small ruminant breeds (Dorper sheep; Australian Boer goat). Morbidity and mortality rates were significantly (P < 0.001) higher in indigenous Kajli sheep (75.37 and 32.80%) and Beetal goats (81.10 and 37.24%) as compared to Dorper sheep (6.99 and 1.48%) and Australian Boer goat (5.01 and 2.23%). Affected animals exhibited high fever, severe diarrhea, abdominal pain, respiratory distress and nodular lesions on lips and nostrils. Thick mucous discharge was oozing out from nostrils. On necropsy, lungs were congested and pneumonic, with nodular and cystic appearance. Intestines were hemorrhagic with zebra stripping. Characteristic histopathological lesions of PPR were noted in intestines, lymphoid organs and lungs. In GI tract, stunting and blunting of villi, necrotic enteritis, and infiltration of mononuclear cells in duodenum, jejunum and ileum. Small intestines exhibited diffuse edema of the submucosa along with proliferation of fibrocytes leading to thickened submucosa which has not been reported previously. Lymphoid organs showed partial to complete destruction of lymphoid follicles. Lesions of the respiratory tract included depictive of bronchopneumonia, severe congestion of trachea and apical lobe of lungs with deposition of fibrinous materials. Histopathological lesions of respiratory tract were severe and characteristic of broncho-interstitial pneumonia, bronchopneumonia, interstitial pneumonia and fibrinous pneumonia. The alveoli were filled with edematous fluid mixed with fibrinous exudate, numerous alveolar macrophages, mononuclear cells along with thickened interalveolar septa and presence of intranuclear eosinophilic inclusion bodies. One-Step RT-PCR using NP3 and NP4 primers confirmed a PPR virus of 352 bp size in spleen, lungs and mesenteric and brachial lymph node samples. It was concluded that morbidity and mortality due to PPR were significantly higher in indigenous breeds of sheep and goat as compared to imported sheep and goat breeds. PPR has rendered various lesions in GI and respiratory tract which are characteristic in nature for the diagnosis of the disease under field condition.

Patho-bacteriological investigation of an outbreak of Mycoplasma bovis infection in calves - Emerging stealth assault.

Mycoplasma bovis (M. bovis) is an important bacterium, causing severe respiratory infection, and arthritis in dairy animals worldwide. This study is based on 50 suckling calves among which 15 showed respiratory distress, lameness and swollen joints and died later. M. bovis was isolated and identified from all dead (n = 15) and live (17.14%; 06 out of 35) calves on the basis of bacteriological examination. In morbid calves, the carpus and stifle joints were severely affected, while necropsy revealed multiple well-circumscribed calcified abscesses and caseous exudates in cranio-ventral and diaphragmatic lobes of lungs. Suppurative polyarthritis, fibrino-suppurative, teno-synovitis and otitis media were the common and striking lesion observed at postmortem examination. Histopathological examination revealed broncho-interstitial pneumonia and necrotic fibrino-purulent broncho-pneumonia in lungs. Similarly, synovial membranes and joints revealed presence of multiple foci of liquefactive necrosis surrounded by lymphocytes, plasma cells, macrophages and peripheral fibroplasia. In the bacteriological investigations, the characteristic fried egg colonies of M. bovis further confirmed this infection in all suspected cases. In conclusion, the current clinico-histo-pathological features are the depictive picture, and is the first report of M. bovis infection in calves in Pakistan.

In vivo anticoccidial effects of Beta vulgaris (sugar beet) in broiler chickens.

Recently, use of botanicals as an alternative to anticoccidial drugs has been appealing approach for controlling avian coccidiosis. Therefore, this study was conducted to evaluate the anticoccidial activity of aqueous methanolic extract (100, 200 and 300 mg/kg of body weight) of Beta vulgaris (roots) in broiler chicks. A total of 315 day old broiler chicks were divided into seven equal groups (A, B, C, D, E, F and G). At 14th day of age, all groups except group G, which served as non infected non medicated control, were infected orally with 60,000 sporulated oocysts of mixed Eimeria species. At the same day, groups A, B and C were treated with graded oral doses of B. vulgaris aqueous methanolic extract (100, 200 and 300 mg/kg of body weight, respectively). Group D was treated with Vitamin-E, group E served as infected medicated control group (Baycox® treated) and group F served as infected non medicated control group (PBS treated). Treatment with extract, reference drug Baycox®, Vitamin E and PBS was continued for three consecutive days (14-16 days of age). Though, not at par with reference drug (Baycox®), B. vulgaris demonstrated good anticoccidial activity adjudged based on considered criteria, i.e., feed conversion ratio, lesion score, oocyst score and oocysts per gram of feces. Results of serum profile of infected chicks revealed no adverse effects of aqueous methanolic extract of B. vulgaris on the experimental chicks.

A review on epidemiology, global prevalence and economical losses of fasciolosis in ruminants.

Fasciolosis is an important plant borne trematode zoonosis in ruminants caused by the Fasciola hepatica and Fasciola gigentica, It is classified as a neglected tropical disease and found in more than 50 countries especially where sheep and cattle are reared. Fasciolosis is a serious animal health problems in many rural and urban areas of world, causing significant financial losses due to decrease in production and viscera condemnation in animals. Accurate diagnosis of fasciolosis is always remained a challenging task for the field practitioners. There is no comprehensive summary on the occurrence and distribution of the infection at international level. Therefore, we intended to provide a complete overview on the prevalence and epidemiology of fasciolosis in farm animals from a global prospective. It includes to map the global distribution of fasciolosis in different areas of the world to identify the endemic regions which may be a source of potential disease outbreak. The financial liability related to fasciolosis on the livestock production has also been addressed. For this purpose, the published data during 2000-2015 (15 years) on fasciolosis was reviewed and collected by electronic literature search of four databases including Google, PubMed, Science Direct, and Web of Science. Data presented are contemplated to enhance our current understanding of the parasite's geographical distribution, host range, and economic losses. Information provided would be useful for the application of more effective control strategies against fasciolosis in different geo-economics regions of the world.

Zinc Oxide Nanoparticles Significant Role in Poultry and Novel Toxicological Mechanisms.

Zinc oxide nanoparticles (ZnO NPs) have involved a lot of consideration owing to their distinctive features. The ZnO NPs can be described as particularly synthesized mineral salts via nanotechnology, varying in size from 1 to 100 nm, while zinc oxide (ZnO), it is an inorganic substrate of zinc (Zn). The Zn is a critical trace element necessary for various biological and physiological processes in the body. Studies have revealed ZnO NPs' efficient immuno-modulatory, growth-promoting, and antimicrobial properties in poultry birds. They offer increased bioavailability as compared to their traditional sources, producing better results in terms of productivity and welfare and consequently reducing ecological harm in the poultry sector. However, they have also been reported for their toxicological effects, which are size, shape, concentration, and exposure route dependent. The investigations done so far have yielded inconsistent results, therefore, a lot of additional studies and research are required to clarify the harmful consequences of ZnO NPs and to bring them to a logical end. This review explores an overview of efficient possible role of ZnO NPs, while comparing them with other nutritional Zn sources, in the poultry industry, primarily as dietary supplements that effect the growth, health, and performance of the birds. In addition to the anti-bacterial mechanisms of ZnO NPs and their promising role as antifungal, and anti-colloidal agent, this paper also covers the toxicological mechanisms of ZnO NPs and their consequent toxicological hazards to vital organs and the reproductive system of poultry birds.

Role of Silver Nanoparticles for the Control of Anthelmintic Resistance in Small and Large Ruminants.

Helminths are considered a significant threat to the livestock industry, as they cause substantial economic losses in small and large ruminant farming. Their morbidity and mortality rates are also increasing day by day as they have zoonotic importance. Anthelmintic drugs have been used for controlling these parasites; unfortunately, due to the development of resistance of these drugs in helminths (parasites), especially in three major classes like benzimidazoles, nicotinic agonists, and macrocyclic lactones, their use is becoming very low. Although new anthelmintics are being developed, the process is time-consuming and costly. As a result, nanoparticles are being explored as an alternative to anthelmintics. Nanoparticles enhance drug effectiveness, drug delivery, and target specificity and have no resistance against parasites. Different types of nanoparticles are used, such as organic (chitosan) and inorganic (gold, silver, zinc oxide, iron oxide, and nickel oxide). One of them, silver nanoparticles (AgNPs), has unique properties in various fields, especially parasitology. AgNPs are synthesized from three primary methods: physical, chemical, and biological. Their primary mechanism of action is causing stress through the production of ROS that destroys cells, organs, proteins, and DNA parasites. The present review is about AgNPs, their mode of action, and their role in controlling anthelmintic resistance against small and large ruminants.

Distribution pattern of Crimean-Congo Hemorrhagic Fever in Asia and the Middle East.

Crimean-Congo Hemorrhagic Fever (CCHF) is one of the most important vector-borne diseases of zoonotic potential that can be acquired following the bite of the Hyalomma species of ticks. It is a highly prevalent disease in Asia and the Middle East. The risk factors of this disease are contact with infected tissue, blood, patient, or livestock in the acute viremic phase, infected tick bites, or the manual removal of ticks. The disease is clinically described as progressive hemorrhages, fever, and pain in musculature. Biochemical tests reveal elevated levels of creatinine phosphokinase, alanine transaminase, aspartate aminotransferase, and lactate dehydrogenase. Clotting time is prolonged in pro-thrombin tests, and pathogenesis is mostly related to the disruption of the epithelium during viral replication and indirectly by secreting cytotoxic molecules. These molecules cause endothelial activation and result in the loss of function. Supportive therapy is given through blood or plasma infusions to treat or manage the patients. According to the most advanced studies, CCHF can be treated by Ribavirin, which is an antiviral drug that shows excellent results in preventing the disease. Health-care staff are more prone to infection. The hemorrhagic phase represents a high risk for accidental exposures. This literature review presents a comprehensive overview of the viral epidemiology, zoonotic perspectives, and significant risk factors of CCHF in various Middle East and Asian countries. Furthermore, the pathophysiology and preventive strategies of CCHF have also been discussed as well as legislation and policies regarding public outreach programs, research, and development aimed at infection prevention and control that are required at a global level.

Molecular evidence of toxoplasmosis in people living with HIV in Pakistan.

BACKGROUND: Toxoplasma gondii is a zoonotically important parasite infecting almost all vertebrates. The parasite can easily infect humans through the fecal-oral route, particularly immunocompromised individuals, and children. Due to lack of epidemiological data regarding the occurrence of Toxoplasma gondii in people living with HIV in Pakistan, the present study was designed to estimate the prevalence of Toxoplasma gondii in people living with HIV. METHODS: Briefly, 384 blood samples were randomly collected from people living with HIV registered under Punjab AIDS Control Program. DNA of all samples was extracted and the 18S rRNA gene of T. gondii was amplified through polymerase chain reaction (PCR) by using genus-specific primers. The amplicons were spread on 1.5% agar rose gel and visualized under gel documentation system. Then the amplicons were purified from the gel and subjected to sequence and phylogenetic analysis. RESULTS: Overall prevalence of T. gondii was observed as 23.96%. The prevalence of T. gondii was significantly (p <0.05) higher in females and in middle age group (26-40 years). Similarly, the prevalence was significantly (p <0.05) higher in people with poor socioeconomic status, symptomatic patients, drugs users and people using unfiltered water for drinking. Furthermore, bodyweight (46-70 kg) and dry seasons also significantly (p <0.05) affected the prevalence of T. gondii. CONCLUSIONS: Our findings illustrated a high infection rate of toxoplasmosis in people living with HIV which could be a significant cause of mortality. More studies need to be conducted for other opportunistic infections caused by parasites and other pathogens.

Molecular evidence of Ehrlichia canis, associated risk factors and hematobiochemical analysis in client owned and shelter cats of Pakistan.

Ehrlichiosis is an infectious disease caused by Ehrlichia canis (E. canis) genus and arthropod vectors. It is considered endemic in many parts of the world among dogs. But due to lack of research on cats, there isn't enough information available. The limited reports available on feline Ehrlichiosis relied on the detection of morulae in leukocytes. The current study was designed to detect the molecular prevalence of E. canis in cats along with associated risk factors and hematological analysis. A total of 384 blood samples from cats were collected from various veterinary hospitals and shelter homes and tested by microscopy and Polymerase Chain Reaction (PCR) to identify E. canis. The prevalence of E. canis has been reported at 5/384 (1.30%) and (14/384) 3.65% in cats through microscopy and PCR respectively. DNA sequences revealed significant resemblance with each other and variable resemblance with other Ehrlichia spp. sequences of different species from various countries already deposited on NCBI. Moreover, hematobiochemical and risk factor analysis were also carried out revealing significant results. This study reports first molecular detection of E. canis in client-owned and sheltered cats located in District Lahore, Punjab, Pakistan. Further studies should be conducted to identify its occurrence in the feline population of Pakistan so that control and prevention strategies must be planned accordingly. Due to the zoonotic impact of this pathogen and in perspective of one health, endemic regions of the disease should be identified and possible control measures should be implemented in these regions to minimize the spread of disease to non-endemic regions of the world and from animals to humans.

Application of Cypermethrin-Coated ZnS and ZnO Nanoparticles against Rhipicephalus Ticks.

Rhipicephalus ticks are described as important ticks impacting the costs of livestock rearing and by-products sale. The prevalence and response of ticks towards cypermethrin sprays indicate the need to implement the rational use of acaricides. In our previous studies, ZnO nanoparticles were shown to inhibit the major life-cycle stages of Hyalomma ticks, indicative of promising application of nanomaterials against the hard ticks. The current study was designed to probe into one of alternative options to curtail Rhipicephalus ticks by employing cypermethrin-coated nanoparticles of ZnO (C-ZnO NPs) and ZnS (C-ZnS NPs). The nanocomposites showed a roughly spherical type of morphology and various size dimensions upon characterization using SEM and EDX. Female ovipositioning was declined up to only 48% in ZnS and up to 32% in ZnO NPs even after 28 days in vitro. Similarly, the larval hatching was also impacted, leading to a hatching percentage of 21% and 15% by application of C-ZnS NPs and C-ZnO NPs, respectively. The LC90 in female adult groups were 3.94 mg/L and 4.27 mg/L for the C-ZnO NPs and C-ZnS NPs groups, respectively. Similarly, the larval groups had LC90 of 8.63 and 8.95 mg/L for the C-ZnO NPs and C-ZnS NPs groups. The study is a proof of the concept for incorporating effective and safe nanocomposites as acaricides. The studies on the efficacy and spectrum of non-target effects of nanomaterial-based acaricides can further refine the research on finding novel alternatives for tick control.