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.

Risk analysis and seroprevalence of bovine ephemeral fever virus in Punjab, Pakistan.

Bovine ephemeral fever (BEF) is a vector-borne viral disease caused by the RNA virus which belongs to the genus Ephemerovirus and the family Rhabdoviridae. To evaluate the effect of the risk factors like the breed of cattle and buffaloes, age, sex, lactation, housing and region on the bovine ephemeral fever virus (BEFV) prevalence, ELISA and virus neutralisation (VN) tests (n = 600) were performed for the BEFV prevalence. The seroprevalence in cattle was 45.6% and 42% by ELISA and VN, respectively (P = 0.001). The breed-wise seropositive ratio was (55-64%) in cattle and (22.5-18.3%) in buffaloes by VN and ELISA. The sex-wise prevalence was (40-49.4%) in females and (35.8-46%) in males by VN and ELISA in cattle and a similar prevalence was reported in buffaloes. The age-wise prevalence in bovines by ELISA was 5.33, 22.66 and 17.66% in the age group < 1 year, 1-3 years and > 3 years, respectively. The disease prevalence was higher in the age group of 1-3 years. The prevalence was higher during the 3rd lactation in bovines. The region-wise prevalence was higher in the 07 districts while lower (18-21%) in Rawalpindi District by VN and ELISA, respectively (P = 0.001). Commercial dairy farms of cattle showed a higher disease prevalence (52% and 44%) than non-commercial farms (38% and 36%) by ELISA and VN, respectively (P = 0.227). Exotic cows showed higher disease prevalence (76.67% and 70%) by ELISA and VN. The mortality in bovines was 5% (7.7% and 2.3%) in the cattle and buffaloes. The case fatality of BEFV in bovines was 12.25%. There was a significant effect of the risk factors like the breed, age, sex, lactation, housing and region on the BEFV prevalence. This is the first comprehensive study of BEFV in Pakistan.

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.

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.

Antimicrobial resistance modulation of MDR E. coli by antibiotic coated ZnO nanoparticles.

We evaluated three types of total six preparations against multidrug resistant E. coli i) three antibiotic coated ZnO nanoparticles (gentamicin coated nanoparticle-GNp; chloramphenicol coated nanoparticles-CNp; and both gentamicin & chloramphenicol coated nanoparticle-GCNp), ii) ZnO nanoparticle alone-Np, and iii) two antibiotics used in single (Gentamicin-G; and Chloramphenicol-C). A total of n = 200 sub-clinically positive mastitic milk samples of bovine origin were processed for isolation of MDR E. coli using microbiological and clinical laboratory & standard institute's protocols. ZnO Nps were prepared from zinc acetate dihydrate (Zn (CH3COO)2. 2H2O), polyethylene glycol (C2nH4n+2On+1), and urea (CH4N2O) by standard chemical protocol. Nps were characterized by XRD and STEM analyses while coating of antibiotics on Nps was confirmed by UV-Visible spectrophotometric analysis. Analysis of variance and student t-test were applied at 5% probability using SPSS version 22 statistical software for inferences on obtained data. There was significantly (p < 0.05) lowest minimum inhibitory concentrations (MICs) and highest zone of inhibitions (ZOIs) in case of GCNp (10.42 ± 4.51 µg/mL & 22.00 ± 1.00 mm) followed by GNp (20.79 ± 8.95 µg/mL & 20.00 ± 1.00 mm) and then CNp (25.96 ± 8.95 µg/mL & 12.33 ± 0.57 mm). Percentage increase in ZOI were expressed as 135.8, 78.43, and 312.76% by GCNp when compared with that of G, C, and Np, respectively. GNp and CNp coated preparations exhibited 114.36 and 275.73% increase in ZOI than to that of G and C, respectively. Similar trend was found in percentage reduction of MICs of preparations. Highest filamentation, indicator of bacterial damage, of E. coli was noted at MIC of GCNp followed by GNp and CNp. The study concluded antibiotic coated ZnO nanoparticles significant candidates modulating antibiotic resistance in MDR E. coli.

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.

Frequency distribution of hard ticks (Acari: Ixodidae) infesting bubaline population of district Toba Tek Singh, Punjab, Pakistan.

The current research was conducted to define the epidemiological parameters related to the prevalence and associated risk factors of tick infestation in buffaloes in the Toba Tek Singh District of central Punjab, Pakistan. The prevalence of ticks on buffaloes was 31.21 % (352/1,128). Among the species of ticks, the prevalence of Hyalomma marginatum (75.56 %; 266/352) was higher (P < 0.05; odd's ratios (OR) = 3.09) than Rhipicephalus microplus (24.44 %; 86/352). Female buffaloes (69.60 %; 245/352) and younger animals (59.09 %; 208/352) were more heavily infested than males (30.40 %; 107/352) and adult animals (40.91 %; 144/352), respectively, whereas breed was not a determinant (P > 0.05). With regard to management and husbandry practices, the prevalence of ticks was higher in animals kept on uncemented flooring (54.55 %; 192/352; OR = 1.90) followed in order by partially cemented (28.69 %; 101/352; OR = 1.71) and fully cemented flooring (16.76 %; 59/352). With regard to feeding systems, grazing animals (64.20 %; 226/352) were more burdened compared to stall-fed animals (35.80 %; 126/352). The highest tick prevalence was recorded in closed housing systems (52.27 %; 184/352), followed by semi-closed (34.09 %; 120/352; OR = 1.53), and open housing systems (13.64 %; 48/352). Rope-tied animals (70.73 %; 249/352) were more parasitized (P > 0.05) than open (29.27 %; 103/352). Prevalence in the study district was highest in tehsil Kamalia followed in order by T.T. Singh and Gojra. The primary body area of infestation by ticks (head, neck, ear, dewlap, back, abdomen, foreleg, shoulder, hind leg, congenital areas, and tail) ranged from highest at inside thigh (17 %) to lowest at rump. In the present survey, the highest prevalence was recorded in July and lowest in December. Comparison of hematological changes showed remarkable differences between infested and non-infested animals, in the form of low values of infested animals, whereas an increment in biochemical parameter values was observed in tick-infested animals. The present study provides significant data to enhance planning for tick control program in the study area.

Protective immunity induced by a DNA vaccine-encoding Toxoplasma gondii microneme protein 11 against acute toxoplasmosis in BALB/c mice.

Toxoplasma gondii is one of the most prevalent intracellular parasites and is threatening the health of both humans and animals, therefore causing incalculable economic losses worldwide. Vaccination is thought to be an efficient way of controlling toxoplasmosis. T. gondii microneme protein 11 (MIC11) is a soluble microneme protein which is presumably considered facilitating the early stage of cell invasion. To evaluate the protective efficacy of T. gondii MIC11, in the present study, a new DNA vaccine-encoding the α-chain of T. gondii MIC11 was constructed using the pcDNA3.1 vector. Expression of MIC11 from this vector was confirmed by indirect immunofluorescence assay following transfection into baby hamster kidney (BHK) cells. Intramuscular immunization of BALB/c mice with pcDNA/MIC11 was carried out to evaluate the immune responses by serum antibodies titers, lymphoproliferation assay, and cytokines assay. The protective efficacy was evaluated by survival rate in mice after challenging with highly virulent strain of T. gondii. The results demonstrated that this vaccination elicited significant humoral responses and T. gondii lysate antigen (TLA)-stimulated lymphoproliferation (p < 0.05). Compared to controls, the pcDNA/MIC11 immunized mice had high production of IFN-γ, IL-12, and IL-2 (p < 0.05), but not IL-4 (p > 0.05), indicating that a predominant Th1 type response was developed. The vaccination also increased the survival rate of immunized mice when they were challenged with a lethal dose of tachyzoites of T. gondii RH strain. These data suggest that T. gondii MIC11 is a reasonable vaccine candidate deserving further studies, and pcDNA/MIC11 is a potential strategy for the control of toxoplasmosis.

The global burden of fasciolosis in domestic animals with an outlook on the contribution of new approaches for diagnosis and control.

Fasciolosis is an economically important disease for livestock, as well as being zoonotic. Recent figures on the prevalence of this disease have caused alarm concerning its potential for an increased prevalence in the future. The prevalence of fascioliosis has been documented from different regions of the world, helping us identify areas where future research needs to be focused. This manuscript is a review of the current status of the disease, the pathogenic species involved, diagnostic techniques (with new modifications and comparative specificity, sensitivity, and rapidity of these tests), chemotherapy, and vaccination. This also encompasses inaccurate reports on vaccination and drug development as well as the latest technologies to find promising candidates for drugs and vaccines. Drugs with lower efficacy have been used on some farms which lead to exacerbation of the clinical disease, presumably due to the development of drug resistance. Future studies should be focused on (1) the use of the most reliable diagnostic tests for periodic monitoring of the disease, (2) insights of the ecobiology and transmission dynamics of the snail intermediate host and the best possible methods of their control, (3) in vitro and in vivo testing of chemotherapeutic compounds using sensitive methods, and (4) the identification of novel drug and vaccine candidates using modern molecular markers. This approach may help increase the reliability of chemotherapeutic agents and control nuisance, ultimately reducing the economic losses attributable to the livestock industry around the world.

Detection of Mycoplasma wenyonii in cattle and transmission vectors by the loop-mediated isothermal amplification (LAMP) assay.

Mycoplasma wenyonii is a wall-less hemotrophic prokaryote with worldwide distribution. This paper describes the development of a LAMP method targeting 16S rRNA for specific detection of M. wenyonii in its vectors and cattle. The LAMP method is specific for M. wenyonii detection and more sensitive than PCR. A total of 330 blood samples from cattle were tested by LAMP and PCR detection, 71 (21.5 %) samples were positive by the LAMP, while only 62 (18.8 %) were positive by PCR. For detecting transmission vectors, 26 lice, 30 flies, and 26 mosquitoes were collected and 18 lice, 20 flies, and 21 mosquitoes were tested positive by LAMP and PCR. These results indicate that the LAMP assay is a simple and convenient diagnostic tool for M. wenyonii detection and can be used in epidemiological surveys.

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.

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.

Risk factors associated with prevalence and major bacterial causes of mastitis in dromedary camels (Camelus dromedarius) under different production systems.

A cross-sectional study was conducted in desert environment of Jhang (Pakistan) from November 2008 to October 2009 on she-camels kept under pastoralist conditions to determine the prevalence of mastitis, impact of risk factors, and isolate the dominant mastitis-causing bacteria on total of 150 lactating she-camels by using clinical examination and surf field mastitis test. From the 150 she-camels examined, 69 (46%) were positive for mastitis at animal level, 12 (8%) clinical, and 57 (38%) subclinical. Age, parity number, stage of lactation, breed, production system, hygiene of milking process, and presence of lesion on udder/teat were found significantly associated (p<0.05) with the prevalence of mastitis in she-camels. There was the lowest prevalence (33.33%; 15 of 45) of mastitis in she-camels of 5-7 years of age, while the highest (80%; 12 of 15) in the animals aged between 14 to 16 years. Stage of lactation significantly affected (p<0.05) and was found to be associated with the prevalence of mastitis being the highest (54.55%; 18 of 33) during the initial stage of lactation (0 to 1 month) followed by last 2 months (10-12 months) as 54.17% and mid-stages (1-3 and 3-10 months) of lactation as 28.57% (6 of 21) and 37.50% (9 of 24), respectively. According to breed of camels, it was noted that the prevalence of mastitis affected significantly (p<0.05) being the highest in crossbred (Desi × Mareecha) as 51.39% (37 of 72) followed in order by Mareecha and Desi as 43.14% and 37.04%, respectively. Staphylococcus (42.19%) and Streptococcus (15.63%) genera were the dominant isolates identified. Good hygiene in milking process, milking clinically infected she-camels at last, culling chronic mastitis carriers, treating clinically infected she-camels, and dry period therapy could reduce the prevalence of contagious mastitis in the study area.

Factors affecting yield and composition of camel milk kept under desert conditions of central Punjab, Pakistan.

This study was planned to study the herd composition, farming system, and reproductive traits and to evaluate the effect of season, stage of lactation and parity on milk production, and composition of camels kept under pastoral environment of central Punjab, Pakistan. Based on purposive sampling method, 50 herds belonging to small, medium, and large-sized herds were selected. From these herds, 1,137 she-camels were entered in this study and their composite milk samples were collected and analyzed through standard procedures to determine the milk yield and percentages of milk contents. The results showed that the male camels constituted a lesser percentage (p < 0.05: 43.08; 380/882) in the total herd composition compared to that of she-camels (56.92; 502/882). The mean daily milk yield was 8.17 ± 0.09 L and mean percentage of fat was 3.79 ± 0.13%, protein was 3.66 ± 0.07%, lactose was 5.15 ± 0.09%, ash was 0.81 ± 0.02%, acidity was 0.20 ± 0.01%, solids not fat (SNF) was 9.63 ± 0.15%, total solids was 13.42 ± 0.21, and moisture was 86.58 ± 0.43. Mean daily milk yield was significantly higher (p < 0.01) during rainy season followed by winter season, warm dry summer, and hot summer season. Milk fat and protein contents were the highest in hot dry summer, while lactose contents were higher during rainy season. The stage of lactation and parity confirmed to impinge significantly (p < 0.01) on protein, lactose, and SNF. The present study will be helpful to design measures for the eradication of reproductive constraints and for the improvement of milk yield and composition in order to make camel rearing an economical proposition.

Nanoparticles: Synthesis and Their Role as Potential Drug Candidates for the Treatment of Parasitic Diseases.

Protozoa, helminths and ectoparasites are the major groups of parasites distributed worldwide. Currently, these parasites are treated with chemotherapeutic antiprotozoal drugs, anti-helminthic and anti-ectoparasitic agents, but, with the passage of time, resistance to these drugs has developed due to overuse. In this scenario, nanoparticles are proving to be a major breakthrough in the treatment and control of parasitic diseases. In the last decade, there has been enormous development in the field of nanomedicine for parasitic control. Gold and silver nanoparticles have shown promising results in the treatments of various types of parasitic infections. These nanoparticles are synthesized through the use of various conventional and molecular technologies and have shown great efficacy. They work in different ways, that include damaging the parasite membrane, DNA (Deoxyribonucleic acid) disruption, protein synthesis inhibition and free-radical formation. These agents are effective against intracellular parasites as well. Other nanoparticles, such as iron, nickel, zinc and platinum, have also shown good results in the treatment and control of parasitic infections. It is hoped that this research subject will become the future of modern drug development. This review summarizes the methods that are used to synthesize nanoparticles and their possible mechanisms of action against parasites.

Molecular Epidemiology of Cryptosporidium parvum and Giardia lamblia in Different Water Bodies, Soil, and Vegetables in Pakistan.

There is a dearth of knowledge regarding transmission of Cryptosporidium (C) and Giardia (G) species through water and water-related sources in Pakistan. To this end, we conducted a study to evaluate the prevalence of these parasites in different water bodies, soil, and mixed raw vegetables in Pakistan. Researchers collected 200 samples from each reservoir including municipal water, sewage water, canal water, raw vegetables, soil of public parks, and soil of grazing areas. Researchers amplified the 18S ribosomal RNA gene of parasites using newly designed genus-specific primers through polymerase chain reaction testing. The sequencing analysis revealed that the obtained sequences belonged to C parvum and G lamblia. Phylogenetic clustering and sequence analysis of C parvum showed that the C parvum Pak1 (OM540369) and C parvum Pak2 (OM540370) as well as C parvum Pak3 (OM510450) and C parvum Pak6 (OM510445) were closely similar to each other. In the case of G lamblia, all the sequences appeared in the same clade. The epidemiological data showed lower prevalence of C parvum (11.5%) in all reservoirs, compared with G lamblia (20.5%). Among different reservoirs, prevalence of Cryptosporidium and Giardia was observed in sewage water (C = 13%, G = 26.5%), municipal water (C = 10%, G = 35%), canal water (C = 9.5%, G = 18.5%), raw vegetables (C = 5.5%, G = 8%), soil of public parks (C = 13%, G = 14%), and soil of grazing areas (C = 18.5%, G = 21.5%). Among targeted risk factors, poor hygienic conditions significantly affected the prevalence of parasites in sewage water (C = 20%, G = 41.2%), municipal water (C = 14.7%, G = 48.8.5%), raw vegetables (C = 11.3%, G = 15.1%), and soil of public parks (C = 19.5%, G = 21.9%). Similarly, contamination with sewage waste significantly affected (P < .05) the prevalence of these parasites in municipal water (C = 13.6%, G = 41.1%), canal water (C = 13.3%, G = 25%), raw vegetables (C = 12.1%, G = 15.1%), and soil of public parks (C = 23.3%, G = 15%). Results of this study illustrated a high risk of parasitic zoonosis through water bodies, soil, and vegetables.

Prevalence, risk factors and first record of mitochondrial cox1 gene-based molecular characterization of Paramphistomum epiclitum from Pakistan.

Parasitic infestations are one of the major threats to the livestock industry in Pakistan. These have a negative impact on the production of domesticated livestock species. Paramphistomes belong to the superfamily Paramphistomoidea and are involved in infecting ruminants all over the world. To date, there was no information on mitochondrial DNA-based molecular characterization of Paramphistomum epiclitum from Pakistan. To close this research gap, this study was designed to provide insights into the epidemiology of Paramphistomum species. Paramphistomum epiclitum isolates were recovered from the rumen of small ruminants slaughtered at an abattoir located in Faisalabad city and animal demographics were recorded. DNA was extracted and mitochondrial cox1 was amplified and sequenced. Prevalence was calculated along with a 95% confidence interval in various groups. The chi-square test was applied to determine the association between different variables under investigation. A phylogenetic tree was constructed based on the Bayesian method. Population diversity indices were calculated using DnaSP 4.5 software. A total of 43 mutations were observed among 7 haplotypes. Negative values of Fu's Fs values, and Tajima's D indicated population expansion. Deworming, season, and grazing were the variables that significantly correlate (p < 0.05) with the prevalence of P. epiclitum. The high prevalence of P. epiclitum demonstrates that more studies are indeed needed to further understand the prevalence and distribution of P. epiclitum in definitive and all potential intermediate hosts in addition to intraspecies variation and relationship with populations from other locations.

Resistance to Cypermethrin Is Widespread in Cattle Ticks (Rhipicephalus microplus) in the Province of Punjab, Pakistan: In Vitro Diagnosis of Acaricide Resistance.

Control of the cattle tick Rhipicephalus (R.) microplus mainly relies on chemical acaricides and cypermethrin is the most widely used acaricide in Pakistan. Farmers frequently complain about its low efficacy, thus, the present study was designed to quantify the frequency of cypermethrin resistance in cattle ticks. Engorged female R. microplus were collected and tested for the efficacy of cypermethrin using the FAO-recommended larval packet test. Resistance factors (RF) were estimated at both the lethal concentration for 50% (LC50) and 99% (LC99) of ticks. Thirty-three samples were tested, of which 8/33 (24.24%) were classified as resistant based on the RF50, and all 33 were classified as resistant based on the RF99. In District Sargodha, when only the RF50 was considered, 45.5% of samples were classified as resistant, but at RF99, all tested samples were identified as resistant. In District Okara, the variation in RF50 estimates was 2.2-8.3 and variation in RF99 estimates was 10.6-1139.8. Similar results were found in District Attock, where variations in RF50 were 0.8-8.5 and RF99 ranged from 9-237.3. The study showed that cypermethrin resistance is prevalent in these three districts of Pakistan and is likely to be overestimated by classification based on the RF99.

A new insight in immunomodulatory impact of botanicals in treating avian coccidiosis.

Avian coccidiosis is caused by genus Eimeria (E.) i.e. E. maxima, E. necatrix, E. tenella, E. acervulina, E. brunette and E. mitis and lead to three billion US dollar per year economic loss in poultry industry and reduces the growth performance of birds. To purge undesirable foreign agents, immune system produces a variety of molecules and cells that ultimately neutralize target particles in healthy organisms. However; when this particular system compromises, infection develops and the load of pathogens along with their virulence factors overcome both; innate and adaptive immune systems. Livestock and poultry sectors are important part of agriculture industry worldwide. Due to excessive use of chemotherapeutic agents, pathogens have developed resistance against these agents leading to the great economic losses. Numerous therapeutic approaches are in routine process for the treatment and prevention of various ailments but irrational use of antibiotics/chemicals has raised alarming concerns, like the development of drug resistant strains, residual effects in ultimate users and environmental pollution. These problems have led to the development of alternatives. In this regard, anticoccidial vaccine can be used as an alternative but due to high cost of production, plant derived biological response modifiers and antioxidants compounds are considered as a promising alternative. This review summarizes the immunotherapeutic effects of different compounds particularly with reference to avian coccidiosis.

Prevalence, associated determinants, and in vivo chemotherapeutic control of hard ticks (Acari: Ixodidae) infesting domestic goats (Capra hircus) of lower Punjab, Pakistan.

A total of 800 goats of various breeds, age, and sex were randomly selected from Muzaffargarh (M. garh) and Layyah districts of lower Punjab, Pakistan. The selected goats were visited twice a month to collect information about determinants influencing goat tick infestation prevalence. For acaricidal efficacy, 360 tick-infested adult goats were subjected to an acaricidal treatment and post-treatment quantitative assessment of tick burden. Quantification of adult tick detachment 24 h post-treatment and the duration of treatment efficacy were calculated. Overall prevalence of goat tick infestation in both study districts was 60.1% (481/800). The prevalence was higher in district M. garh than in district Layyah. Tehsil-wise prevalence in district Layyah was highest in tehsil Layyah followed in order by Chaubara and Karor. In district M. garh, highest prevalence was found in tehsil M. garh followed by Kot Addu, Alipur, and Jatoi. Hyalomma a. anatolicum (75.9%; 365/481) and Rhipicephalus sanguineus (24.1%; 116/481) were the predominant species in both the districts. The highest month-wise prevalence was 56.9% and 62.7% in Layyah and M. garh districts, respectively, during July 2008, and the minimum (0%) prevalence was reported in November and December, respectively. Regarding host determinants, female goats were more heavily infested (72.8%) than males (47.5%), and younger animals were (63.5%) more burdened than older ones (56.7%). Teddy goats were the most susceptible breeds followed in order by Beetal, cross-bred, Nachi, and Dera Din Pannah. The preferred sites of attachment were inside and outside of the ear. Both the ivermectin (IVM)- and cypermethrin (CYM)-treated groups resulted in significantly lower (P < 0.05) tick counts relative to controls on all post-treatment counting days. The lowest tick burden in the IVM-treated group was significantly higher (P < 0.05) as compared to the CYM-treated group, the latter being close to zero. Hence, the in vivo efficacy trials of injectable IVM vs CYM pour-on revealed better results for the latter. These observations provided the first insights into what determinants impact goat tick infestation, and laid a foundation for planning of future control programs in the lower Punjab, Pakistan.