Development of egg yolk-based polyclonal antibodies and immunoprophylactic potential of antigen-antibody complex against infectious bursal disease.

A study conducted in the Faisalabad district sampled 50 cases across five IBD outbreaks, revealing an alarming 80 % infection rate among poultry. Our research focused on developing an immune complex (Antigen-antibody complex) with potential immunoprophylactic benefits to counter this formidable threat. Our study was based on producing egg yolk-derived polyclonal antibodies (IgY) targeting IBDV. Commercial layer birds were immunized with inactivated IBDV, yielding IgY antibodies extracted from their eggs exhibited substantially higher and more enduring antibody titers, with a geometric mean titer of 104. Further research involved the creation of an immune complex (ICx) where antigen was extracted from infected bursae tissues. The immunogenic response of ICx was assessed in poultry birds after a challenge with a virulent strain of IBD virus and compared to a conventional IBDV vaccine in poultry. Results revealed significantly higher and more enduring antibody titers induced by the ICx, offering enhanced protective immunity against the IBDV challenge, as evidenced by lower Bursa to bodyweight ratios in vaccinated birds.

Phytochemical analysis and gastroprotective effect of Stellaria media (L.) Vill. methanolic extract on piroxicam-induced gastric ulcer in Wistar rats.

Stellaria media L. has traditionally been used to treat inflammatory and gastrointestinal ailments. This study aimed to phytochemically characterize the S. media extract and explore its anti-ulcer efficacy against piroxicam-induced stomach lesions in Wistar rats. Phytochemical analysis was performed and antioxidant capacity of extract was determined using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. In vivo, piroxicam (30mg/kg) was administered to induce gastric ulceration. Gastro protective effect of S. media extract was observed at 150, 300 and 450mg/kg, respectively. While omeprazole (20mg/kg) was used as a conventional anti-ulcer drug. After oral treatment for 14 days, stomach acidic secretions, ulcerogenic indices, hematological markers and oxidative stress parameters were assessed along with histological examination. The existence of polyphenol contents in S. media extract was confirmed in correlation to a marked DPPH inhibition (IC50 27.94µg/mL). S. media extract resulted in a dose-dependent elevation in gastric pH while a decrease in acid volume, acidity and ulceration. Also, S. media extract administration restored the impaired hematological markers (RBCs, Hb, WBCs and PLTs) and decreased oxidative stress by reducing oxidants (TOS and MDA) while raising antioxidants (TAC and CAT). Furthermore, gastric histological results corroborated the aforementioned findings. Conclusively, S. media could provide a promising protective effect against drug-induced gastric ulceration.

Curcumin Co-Encapsulation Potentiates Anti-Arthritic Efficacy of Meloxicam Biodegradable Nanoparticles in Adjuvant-Induced Arthritis Animal Model.

This study aimed to evaluate the anti-arthritic activity of curcumin and meloxicam co-loaded PLGA nanoparticles in adjuvant-induced arthritic rats. PLGA nanoparticles encapsulating curcumin (nCur) and meloxicam (nMlx) alone and in combination (nCur/Mlx) were used to characterize zeta size and potential, polydispersity index, encapsulation efficiency (%), compound-polymer interactions (FT-IR analysis), and surface morphology (SEM imaging). In vivo, Complete Freund's adjuvant-induced arthritic rats were intraperitoneally (i.p.) administered with curcumin, meloxicam, curcumin plus meloxicam, nCur, nMlx, and nCur/Mlx for 28 consecutive days. Results showed that nCur, nMlx, and nCur/Mlx significantly (p ≤ 0.05) reduced paw swelling and arthritic score, restored body weight and the immune organ index (thymus and spleen), as well as attenuated serum inflammatory markers (RF, CRP, and PGE2) and oxidative stress parameters (MDA, SOD, and CAT) in adjuvant-induced arthritic rats compared to free compounds. In addition, mono- and dual-compound-loaded nanoparticles significantly (p ≤ 0.05) down-regulated pro-inflammatory cytokines (TNF-α, IL-1ß, and IL-6), up-regulated anti-inflammatory cytokines (IL-4, IL-10, and IFN-γ), and modulated OPG and RANKL expressions in paw tissue. The aforementioned results were further confirmed through radiological and histopathological examinations. Furthermore, the anti-arthritic effect of nCur/Mlx was notably (p ≤ 0.05) enhanced compared to nCur or nMlx alone. In conclusion, the co-nanoencapsulation of curcumin could potentiate the anti-arthritic activity of meloxicam and could provide a novel therapeutic approach for the formulation of nanocarrier pharmaceutical products for the management of arthritis.

Model nematodes as a practical innovation to promote high throughput screening of natural products for anthelmintics discovery in South Asia: Current challenges, proposed practical and conceptual solutions.

With the increasing prevalence of anthelmintic resistance in animals recorded globally, and the threat of resistance in human helminths, the need for novel anthelmintic drugs is greater than ever. Most research aimed at discovering novel anthelmintic leads relies on high throughput screening (HTS) of large libraries of synthetic small molecules in industrial and academic settings in developed countries, even though it is the tropical countries that are most plagued by helminth infections. Tropical countries, however, have the advantage of possessing a rich flora that may yield natural products (NP) with promising anthelmintic activity. Focusing on South Asia, which produces one of the world's highest research outputs in NP and NP-based anthelmintic discovery, we find that limited basic research and funding, a lack of awareness of the utility of model organisms, poor industry-academia partnerships and lack of technological innovations greatly limit anthelmintics research in the region. Here we propose that utilizing model organisms including the free-living nematode Caenorhabditis elegans, that can potentially allow rapid target identification of novel anthelmintics, and Oscheius tipulae, a closely related, free-living nematode which is found abundantly in soil in hotter temperatures, could be a much-needed innovation that can enable cost-effective and efficient HTS of NPs for discovering compounds with anthelmintic/antiparasitic potential in South Asia and other tropical regions that historically have devoted limited funding for such research. Additionally, increased collaborations at the national, regional and international level between parasitologists and pharmacologists/ethnobotanists, setting up government-industry-academia partnerships to fund academic research, creating a centralized, regional collection of plant extracts or purified NPs as a dereplication strategy and HTS library, and holding regional C. elegans/O. tipulae-based anthelmintics workshops and conferences to share knowledge and resources regarding model organisms may collectively promote and foster a NP-based anthelmintics landscape in South Asia and beyond.

Anti-Pyretic, Analgesic, and Anti-Inflammatory Activities of Meloxicam and Curcumin Co-Encapsulated PLGA Nanoparticles in Acute Experimental Models.

Herein, we evaluated the in vivo effects of meloxicam and curcumin co-encapsulated PLGA nanoparticles in experimental acute models of pyrexia, nociception, and inflammation. Seven groups (n = 6) were designed for each investigation and pretreated intraperitoneally (i.p.): the control group, meloxicam (4 mg/kg b.w.), curcumin (15 mg/kg b.w.), and equivalent content containing PLGA capped nanoparticles of meloxicam (Mlx-NP) and curcumin (Cur-NP) alone and in combination (Mlx-Cur-NP; at two doses). The results showed that PLGA encapsulation significantly (p ≤ 0.05) improved the in vivo activities of each compound. Furthermore, co-encapsulation of meloxicam and curcumin potentiated the anti-pyretic effect on yeast-induced pyretic rats, anti-nociceptive effect on nociception induced in rats by formalin and heat, and anti-edematogenic activity in xylene-induced ear edema in rats in a dose-dependent manner. In carrageenan-induced paw inflammation in rats, meloxicam and curcumin co-loading (Mlx-Cur-NP) resulted in significant (p ≤ 0.05) inhibition of paw inflammation, reduction in TNF-α and PGE2 levels, downregulation of expressions of pro-inflammatory cytokines (TNF-α, IL-1ß, and IL-6), as well as a decrease in histopathological changes and TNF-α immunoexpression in paw tissues. Moreover, Mlx-Cur-NP demonstrated noteworthy potentiation in pharmacological effects compared to free compounds and mono-compound-loaded nanoparticles. Thus, the association of meloxicam with curcumin in a biodegradable nanocarrier system could provide a promising anti-pyretic, anti-nociceptive, and anti-inflammatory therapeutic approach for acute conditions.

Crimean-Congo hemorrhagic fever: Immunopathogenesis and recent advances in the development of vaccines.

Crimean-Congo hemorrhagic fever is a serious vector-borne zoonotic viral infection which leads to severe illness and fatalities in people living in endemic regions and becoming infected sporadically. Hyalomma ticks are responsible for the transmission of the virus which belongs to the family Nairoviridae. This disease spreads through ticks bite, infected tissues, or blood of viremic animals, and from infected humans to others. Serological studies also indicate the presence of the virus in various domestic and wild animals to be a risk factor for the transmission of the disease. Crimean-Congo hemorrhagic fever virus elicits many immune responses during the infection including inflammatory, innate, and adaptive immune responses. The development of an effective vaccine could be a promising method for the control and prevention of disease in endemic areas. The purpose of this review is to highlight the importance of CCHF, its mode of transmission, the interaction of the virus with the hosts and ticks, immunopathogenesis, and advances in immunization.

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.

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.

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.

Molecular Characterization and Demographic Study on Infectious Bursal Disease Virus in Faisalabad District.

The re-emergence of virulent strains of the Infectious Bursal Disease Virus (IBDV) leads to significant economic losses of poultry industry in Pakistan during last few years. This disease causes the infection of bursa, which leads to major immune losses. A total number of 30 samples from five IBD outbreaks during the period of 2019-20 were collected from different areas of Faisalabad district, Pakistan and assayed by targeting the IBD virus VP2 region through RT-PCR. Among all the outbreaks, almost 80% of poultry birds were found positive for the IBDV. The bursa tissues were collected from the infected birds and histopathological examination of samples revealed severe lymphocytic depletion, infiltration of inflammatory cells, and necrosis of the bursa of Fabricius (BF). Positive samples were subjected to re-isolation and molecular characterization of IBDV. The Pakistan IBDV genes were subjected to DNA sequencing to determine the virus nucleotide sequences. The sequences of 100 Serotype-I IBDVs showing nearest homology were compared and identified with the study sequence. The construction of the phylogenetic tree for nucleotide sequences was accomplished by the neighbor-joining method in MEGA-6 with reference strains. The VP2 segment reassortment of IBDVs carrying segment A were identified as one important type of circulating strains in Pakistan. The findings indicated the molecular features of the Pakistan IBDV strains playing a role in the evolution of new strains of the virus, which will contribute to the vaccine selection and effective prevention of the disease.

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.

Repellent and acaricidal activity of essential oils and their components against Rhipicephalus ticks in cattle.

Ticks, particularly the Rhipicephalus which are the most prevalent and invasive affect 80 % of the cattle population worldwide. Through transmission of pathogens, tick worry and physical damage to the hides, ticks cause economic loss of billions of dollars each year with 1 billion US dollars loss per annum reported only in Latin-America. These losses can be minimized only by successful management of Rhipicephalus ticks. Various strategies like chemical control, vaccination and biological control are aimed at control of Rhipicephalus ticks. There are some serious limitations associated with them like tick resistance, drug toxicity, antigenic variations etc. In contrast to these issues related with chemical tick control, the botanicals particularly the essential oils obtained from aromatic plants of medicinal importance are eco-friendly and non-toxic to most host. In recent years, essential oils-based control of cattle ticks has gained considerable attraction of scientists all over the world as depicted from this review. A comprehensive effort has been made to critically analyze the role of essential oils in controlling Rhipicephalus ticks with particular emphasis on the mode of action of bioactive compounds both as repellents and acaricides. Furthermore, we have pointed out the most important challenges which need to be addressed for development and commercialization of an essential oil based anti-tick product.

Potential of botanical driven essential oils against Haemonchus contortus in small ruminants / Potencial de aceites esenciales botánicos contra Haemonchus contortus en pequeños rumiantes

The livestock sector is continuously facing problems in controlling parasitic diseases especially Haemonchosis due to emergence of anthelminthic resistance and failure in vaccination control programmes. Therefore, to increase milk and meat production and emerging demand of meat free from drug residues development of new alternative approaches are appealing for prevention and control of Haemonchosis in small ruminants. Among alternatives, plants driven essentials oils have shown promising results in control of Haemonchus contortus infection at various concentrations by different assays including egg hatch assay, larval development assay, larval exsheathment assay and adult motility assay. Essential oils are complex mixtures of various impulsive or volatile compounds which have potential to control Haemonchosis. The current study reviews the therapeutic effects of essential oils of plants against Haemonchus contortus and to be used them against Haemonchus contortus for future perspectives.

El sector ganadero enfrenta continuamente problemas para controlar las enfermedades parasitarias, especialmente la hemoncosis, debido a la aparición de resistencia antihelmíntica y al fracaso en los programas de control de vacunación. Por lo tanto, para aumentar la producción de leche y carne, y la demanda emergente de carne libre de residuos de medicamentos, el desarrollo de nuevos enfoques alternativos es atractivo para la prevención y el control de la hemoncosis en pequeños rumiantes. Entre las alternativas, los aceites esenciales producidos por las plantas han mostrado resultados prometedores en el control de la infección por Haemonchus contortus a diversas concentraciones mediante diferentes ensayos, incluido el análisis de eclosión de huevos, el desarrollo de larvas, el análisis de vaciado de larvas y el ensayo de motilidad en adultos. Los aceites esenciales son mezclas complejas de varios compuestos impulsivos o volátiles que tienen potencial para controlar la hemonchosis. Este estudio revisa los efectos terapéuticos de los aceites esenciales de las plantas contra Haemonchus contortus y evalúa sus perspectivas futuras como agentes para combatir las enfermedades causadas por este parásito.

Acaricidal and insecticidal effects of essential oils against ectoparasites of veterinary importance / Efectos acaricidas e insecticidas de los aceites esenciales contra los ectoparásitos de importancia veterinaria

Ectoparasitism in animals has become an issue of great concern that needs to be resolved to prevent huge economic losses occurring to livestock industry all over the world. Synthetic adrugs have been playing a major role in controlling ectoparasites, but their frequent and irrational use has resulted in drug resistance to routinely used chemicals and their residual effects on food and environment. Therefore, this approach of using chemical acaricides and insecticides is losing its popularity and effectiveness in controlling ectoparasites. So, the development of alternative approaches in ectoparasite management is currently required. Among alternative protocols, plants and their essential oils have played remarkable role in controlling different ectoparasites (ticks, flies, mites, lice) of veterinary importance. Essential oils have been proved to be cheaper, more effective and safer therapeautic agents against different ectoparasites of livestock importance.

En los animales el ectoparasitismo se ha convertido en un tema de gran preocupación que debe resolverse para evitar que se produzcan grandes pérdidas económicas para la industria ganadera en todo el mundo. Los aditivos sintéticos han desempeñado un papel importante en el control de los ectoparásitos, pero su uso frecuente e irracional ha dado como resultado la resistencia a los fármacos utilizados habitualmente y efectos residuales sobre los alimentos y el medio ambiente. Por lo tanto, el enfoque basado en el uso de acaricidas e insecticidas químicos está perdiendo popularidad y efectividad en el control de los ectoparásitos. Por lo tanto, actualmente se requiere el desarrollo de enfoques alternativos en el manejo de ectoparásitos. Entre los protocolos alternativos, las plantas y sus aceites esenciales han jugado un papel notable en el control de diferentes ectoparásitos (garrapatas, moscas, ácaros, piojos) de importancia veterinaria. Se ha demostrado que los aceites esenciales son agentes terapéuticos más baratos, más efectivos y más seguros contra diferentes ectoparásitos de importancia ganadera.

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.

Prevalence and associated risk factors of haemoparasites, and their effects on hematological profile in domesticated chickens in District Layyah, Punjab, Pakistan.

Several haemoparasites commonly infect avian species, including chicken that significantly decline productivity and ultimately lead to high mortality rate. The current study was designed to assess the prevalence and associated risk factors of haemoparasites in domesticated chickens in and around District Layyah, Punjab Province, Pakistan. For this purpose, blood samples from a total of 384 backyard chickens were collected from wing vein using sterile insulin syringe. The parasites were identified from Giemsa stained thin blood smears based on morphological features using standard keys. Results demonstrated that a total of 265 backyard chickens (69%) were infected with haemoparasites in District Layyah. According to genus-wise distribution, 31.5%, 24.4% and 13% prevalence of genera Plasmodium/Haemoproteus, Leucocytozoon and mixed species were recorded, respectively. Among associated risk factors, the prevalence was relatively higher in females, chicks, naked neck breeds, scavengers feeding patterns and chickens reared at fully open coops type. The information given in the study could be of much importance in planning of an effective haemoparasites control program at District and Provincial level.

Determination of florfenicol residues in broiler meat and liver samples using RP-HPLC with UV-visible detection.

BACKGROUND: Broilers are vulnerable to various types of microorganisms, including Salmonella, Escherichia coli and Staphylococcus aureus, resulting in multiple infections. Broad-spectrum antibacterial drugs such as florfenicol (FF) are widely used in the treatment of such infections. Suspected residues of these drugs in body tissues of treated birds can be passed to humans through meat consumption and thus lead to serious ill effects on human health. The present study was designed to estimate the presence of FF residues in broiler meat and liver samples. RESULTS: The mean residual concentrations of FF in broiler meat and liver samples were 311.42 ± 186.56 and 2585.44 ± 1759.71 µg kg(-1) respectively, which are higher than their respective maximum residual limits (MRLs). The results showed that 126 and 24 samples were FF-positive and FF-negative respectively. Of the positive samples, 84 and 42 samples were above and below the MRL respectively. CONCLUSION: The results indicate the presence of FF residues in broiler meat and liver samples. Usage of this contaminated meat causes resistance in consumers and poses a public health threat. Thus there is a need to educate farmers about the ill effects of residual drugs on human health and their withdrawal times in poultry. © 2015 Society of Chemical Industry.

Syringe test (modified larval immersion test): a new bioassay for testing acaricidal activity of plant extracts against Rhipicephalus microplus.

We report a new bioassay "syringe test" (modified larval immersion test) for in vitro evaluation of acaricidal activity of crude plant extracts. Prepared syringes, containing eggs of tick, were incubated until 14 d after hatching of eggs, when the bioassay was performed on the larvae. Lethal concentrations for 50% of larvae (LC(50)), LC(90) and LC(99) values were calculated for each tested product. 95% confidence intervals for LC(50) were very narrow, indicating a high degree of repeatability for the new bioassay on larvae of R. microplus. Bioassays were applied to six crude aqueous-methanol extracts from five plants (Acacia nilotica, Buxus papillosa, Fumaria parviflora, Juniperus excelsa, and Operculina turpethum), of which three showed discernible effects. Twenty-four hours post exposure, LC(99) values were 11.9% (w/v) for F. parviflora, 20.8% (w/v) and 29.2% (w/v) for B. papillosa and A. nilotica, respectively. After six days of exposure these values were; 9.1% (w/v), 9.2% (w/v) and 15.5 (w/v) for F. parviflora, A. nilotica and B. papillosa, respectively.

Atypical (RIO) protein kinases from Haemonchus contortus--promise as new targets for nematocidal drugs.

Almost nothing is known about atypical kinases in multicellular organisms, including parasites. Supported by information and data available for the free-living nematode, Caenorhabditis elegans, and other eukaryotes, the present article describes three RIO kinase genes, riok-1, riok-2 and riok-3, from Haemonchus contortus, one of the most important parasitic nematodes of small ruminants. Analyses of these genes and their products predict that they each play critical roles in the developmental pathways of parasitic nematodes. The findings of this review indicate prospects for functional studies of these genes in C. elegans (as a surrogate) and opportunities for the design of a novel class of nematode-specific inhibitors of RIO kinases. The latter aspect is of paramount importance, given the serious problems linked to anthelmintic resistance in parasitic nematode populations of livestock.

Anthelmintic activity of Ziziphus nummularia (bark) and Acacia nilotica (fruit) against Trichostrongylid nematodes of sheep.

ETHNOPHARMACOLOGICAL RELEVANCE: Ziziphus nummularia (Rhamnaceae) and Acacia nilotica (Fabaceae) are being used as anthelmintics in ethnoveterinary medicinal system of Pakistan. AIM OF THE STUDY: Present study was conducted to determine the anthelmintic activity of Ziziphus nummularia (bark) and Acacia nilotica (fruit) in order to justify their traditional use in veterinary medicine. MATERIALS AND METHODS: In vitro anthelmintic activity of crude methanolic extract (CME) of both the plants was determined against Haemonchus contortus by the adult motility assay, the egg hatch test and the larval development assay. In vivo anthelmintic activity was evaluated in sheep naturally infected with gastrointestinal nematodes by administering increasing doses of crude powder (CP) and CME (1.0-3.0 g/kg). RESULTS: Both the plants exhibited dose- and time-dependent anthelmintic effects by causing mortality of worms, and inhibiting egg hatching and larval development. Acacia nilotica (LC(50)=512.86 and 194.98 microg/ml) was found to be more potent than Ziziphus nummularia (LC(50)=676.08 and 398.11 microg/ml) in egg hatch test and larval development assay, respectively. In vivo, maximum fecal egg count reduction (84.7%) was recorded on day 13 post-treatment in sheep treated with Ziziphus nummularia CME (3.0 g/kg) followed by 78.5% on same day with Acacia nilotica CME (3.0 g/kg). CONCLUSIONS: These data show that both Ziziphus nummularia and Acacia nilotica possess anthelmintic activity in vitro and in vivo, justifying their use in traditional veterinary medicine in Pakistan.