Garlic and papaya lack control over gastrointestinal nematodes in goats and lambs

Gastrointestinal nematodes (GIN) continue to hinder small ruminant production because of anthelmintic resistance and lack of effective products for GIN control in organic production. The objective of this study was to examine the effectiveness of a commercially available certified organic garlic product, fresh garlic juice, or garlic bulbs as an anthelmintic to control GIN in goats and papaya seeds for GIN control in lambs. In the first experiment, weaned meat goat kids were administered water or a commercial garlic juice product (n = 7/treatment). In the second experiment, kids were administered water, fresh garlic juice, or fed garlic bulbs (n = 9 or 10/treatment). In a third experiment, lambs were administered nothing or papaya seed in water (n = 12/treatment). Naturally infected goats were supplemented with bermudagrass hay and corn/soybean supplement in the first experiment or maintained on bermudagrass pasture in the second experiment or lambs grazed mixed grasses in the third. Blood and fecal samples were collected on Days 0, 7, and 14 after administration of treatment to examine changes in blood packed cell volume (PCV) and fecal egg counts (FEC). PCV and FEC were similar by Day 14 between kids treated with a commercial garlic juice and water in the first experiment. Similarly, PCV was not different among treatment groups in the second experiment. FEC of the garlic juice group was lower than the other two groups on Day 0, but was similar among groups by Day 14. Deworming was required in one goat within each treatment group by Day 7 and in three kids in the garlic juice group and two in the garlic bulb group by Day 14. No changes in PCV or FEC were apparent in response to papaya seed compared with untreated lambs. Based on the alternative plant products used in this study, garlic or papaya seed is not recommended as an aid to control GIN in goats or lambs.     

Duration of Anthelmintic Effect of Three Formulations of Ivermectin (Oral,Injectable and Pour-on) Against Multiple Anthelmintic-Resistant <Emphasis Type="Italic">Haemonchus contortus</Emphasis> in Sheep

We report the results of investigations that were conducted in a sheep flock in Uttaranchal, India where repeated failure of anthelmintic medication was noted. The study revealed that Haemonchus contortus in sheep had developed resistance to benzimidazoles (fenbendazole, mebendazole and albendazole), imidazothiazole (levamisole) and salicylanide (rafoxanide), while it was fully susceptible to avermectins (ivermectin). Further, the suppression of nematode egg output in faeces of sheep naturally infected with multiple anthelmintic-resistant H. contortus following treatment with ivermectin tablet (0.4 mg/kg body weight (bw), orally), ivermectin injection (1% w/v, 0.2 mg/kg bw, subcutaneously) and ivermectin pour-on (0.5 w/v, 0.5 mg/kg bw) was also studied over a period of 10 weeks post treatment. It was noted that ivermectin tablet after initial clearance of infection (faecal egg count reduction 100%), could not prevent establishment of new patent natural infection for even a single day, while ivermectin pour-on and injection prevented the establishment of new infection for 7 and 14 days post treatment, respectively. Maximum protection period (duration for which mean faecal egg count of sheep reaches 500 eggs per gram of faeces or more) of 68 days was recorded in sheep treated with injectable ivermectin, followed by pour-on (60 days) and oral (53 days) preparations.     

Comparison of two versions of larval development test to detect anthelmintic resistance in Haemonchus contortus

Larval development (LDT) and micro-agar larval development tests (MALDT) were used to compare the reliability and sensitivity of two methods for detecting anthelmintic resistance in Haemonchus contortus. The tests were conducted using three resistant and four susceptible isolates of H. contortus. Both versions of the tests provided comparable results with regard to the characterization of benzimidazole and levamisole susceptibility but neither test was sufficiently sensitive to discrimination between an ivermectin (IVM) susceptible and an IVM resistant isolate. Each test has its own merits with the LDT having the advantage of being less time-consuming.     

Sheep helminth parasitic disease in south eastern Scotland arising as a possible consequence of climate change

The climate in the UK is changing, with a trend towards increased rainfall in the autumn and winter and warmer average temperatures throughout the year. There has also been a 4-week extension of the herbage growing season over the past 40 years. These changes may have implications for the epidemiology of sheep helminth parasites. Here, we describe production-limiting disease outbreaks caused by Haemonchus contortus, Nematodirus battus, Teladorsagia circumcincta and Fasciola hepatica in sheep flocks in south eastern Scotland. The occurrence and timing of these disease outbreaks could not have been predicted in this region highlighting changes in the epidemiology of helminth infections from the patterns historically described. These cases are used to introduce discussion regarding the potential effects of climate change on the epidemiology of helminth parasites and the implications for sheep farming in the UK.     

Anthelmintic efficacy of neem (Azadirachta indica A. Juss) and the homeopathic product Fator Vermes in Morada Nova sheep

Gastrointestinal nematodes are becoming increasingly resistant to the commercial products used to control them. The cost of routine vermifuge applications on herds and the problem of residues in animal products and the environment have prompted research on the anthelmintic activity of plant extracts. This work examines the anthelmintic action of neem and the homeopathic product Fator Vermes in sheep kept in a pasture for 18 months. Forty sheep of the Morada Nova breed were divided into four treatments and the control, according to the EPG. During the experiment, each animal received 100 g/day of shredded corn and did not receive protein supplementation. In treatment 1 (control), the animals received only shredded corn. Treatment 2 received 1.6 g/(animal day) of the homeopathic product mixed with the shredded corn, and treatments 3, 4 and 5 received, respectively, 12.5, 25.0 and 37.5 g/(animal day) of dried Azadirachta indica leaves mixed with the shredded corn. The neem was administered for alternating 15-day periods and the homeopathic product daily for 18 months. There were 39 fortnightly fecal collections made to count the EPG, and fecal cultures were performed monthly. The following genera, in percentage, were identified: Haemonchus: 65.58+/-3.27, Trichostrongylus: 15.92+/-7.38 and Oesophagostomum: 18.50+/-6.22. The treatments evaluated were not effective in controlling gastrointestinal nematodes (P>0.05), whose mean log(10) counts (EPG +1) and standard errors for treatments 1-5 were respectively 3.55+/-0.28; 3.48+/-0.31; 3.90+/-0.29; 2.78+/-0.29 and 3.48+/-0.30. A significant effect (P<0.0001) was observed of the periods of the year when the 39 collections occurred. Because of the diet deficient in raw protein, the sheep had higher average EPG counts, for all the treatments, at the end of the dry season, and the opposite occurred in the middle of the rainy season.     

Green tea flavan-3-ols and oligomeric proanthocyanidins inhibit the motility of infective larvae of Teladorsagia circumcincta and Trichostrongylus colubriformis in vitro

The effects of a hot water infusion and an aqueous acetone extract of green tea (Camellia sinensis) on the motility of infective larvae of the sheep nematodes Teladorsagia circumcincta and Trichostrongylus colubriformis were investigated under in vitro conditions. The infusion and extract dose-dependently inactivated the infective larvae as assessed by the larval migration inhibition (LMI) assay. To determine the components responsible for the inhibitory activity, the hot water infusion and aqueous acetone extract of green tea were fractionated on Sephadex LH-20 and the green tea extract fractions (GTE-I-VIII) characterised by mass spectrometry. The larvae were exposed to increasing concentrations of these GTE fractions. Fractions containing epigallocatechin gallate (EGCG) and proanthocyanidin oligomers were most effective. GTE fractions were more effective against T. circumcincta than T. colubriformis larvae as assessed by the LMI assay.     

Evidence for multiple anthelmintic resistance in sheep and goats reared under the same management in coastal Kenya

Four experiments, two with sheep and two with goats, were carried out to determine the efficacy of ivermectin, fenbendazole, levamisole, closantel and some of their combinations by faecal egg count reduction tests. In the first experiment, injectable ivermectin, oral ivermectin, fenbendazole and levamisole were tested in 6-month-old lambs, and their reduction percentages were 77%, 13%, 42% and 92%, respectively. In the second experiment, with yearling sheep, the reduction percentages were 35% for injectable ivermectin, 32% for fenbendazole, 99% for levamisole, 48% for closantel, 92% for injectable ivermectin combined with fenbendazole, 99% for injectable ivermectin combined with levamisole, and 100% for fenbendazole combined with levamisole. In the study with 18-month-old goats given the same dose rates as those recommended for sheep, the reduction percentages were 73% for injectable ivermectin, 25% for fenbendazole, and 78% for levamisole. Another group of 14-month-old goats was treated with dose rates 1.5 times those recommended for sheep and the reduction percentages were 93% for levamisole, 92% for injectable ivermectin, and 97% for a combination of levamisole and ivermectin. In all experiments with sheep and goats the gastrointestinal nematode parasites identified by larval cultures were Haemonchus contortus, Trichostrongylus spp. and Oesophagostomum spp. The gastrointestinal nematodes of both sheep and goats on this farm are resistant to ivermectin and fenbendazole, whereas levamisole is still effective in sheep, but not in goats. The results are discussed in relation to the farm as a source of breeding stock to smallholder farmers and its potential to spread anthelmintic resistance.     

The effect of long-term feeding of fresh and ensiled cassava (Manihot esculenta) foliage on gastrointestinal nematode infections in goats

The benefit of long-term feeding of fresh or ensiled cassava foliage on gastrointestinal parasite in goats was evaluated. Eighteen male goats (15.15 ± 2.83kg and between 4–6months) were randomly allocated into three treatments supplemented with 200g of wheat bran head⁻¹ day⁻¹. All groups were fed ad-libitum on either grass (CO), fresh cassava (CaF) or ensiled cassava foliage (CaS). At the beginning of the trial, each goat was inoculated with 3000 L3 containing ≈50% Haemonchus contortus. Individual LWt, FEC and PCV were measured at weekly intervals for 10weeks. At the termination of the experiment all goats were slaughtered for worm recovery and enumeration. The goats in CaF and CaS had similar weight gains while those in CO lost weight (p < 0.05) through the trial. FEC in CaF and CaS were lower (p < 0.05) than CO during the patency of parasite infections, but there was no difference between CaF and CaS goats. PCV of all groups decreased from above 30% to around 25% at the end of the trial. The compositions of established worm burdens were mainly H. contortus (19–40%) and Trichostrongylus colubriformis (55–76%). TWB did not differ among the groups, however, CaS significantly reduced H. contortus burdens, as compared to CaF and CO (p ≤ 0.005). Thus, ensiled cassava foliage reduced the H. contortus population while the fresh foliage only reduced worm fecundity.     

Comparative efficacy of pour-on and subcutaneous injection of ivermectin on Melophagus ovinus (L.) in Darab ecotype goats of Southern Iran

The efficacy of ivermectin was evaluated against Melophagus ovinus in Darab ecotype goats of Iran. Twenty-four healthy Iranian crossbreed male goats were randomly divided into three equal groups (n = 8). An experimental infestation was induced in all animals of the three groups with 100 M. ovinus on the body of each animal. Groups 1 and 2 were treated with 1% ivermectin solution at a dosage of 0.5 mg/kg of body weight applied as a pour-on along the dorsal midline and 0.2 mg/kg subcutaneously, respectively; while group 3 was kept as control group. Seven days after infestation ivermectin was administered then the goats were observed for a period of 7 days. Body surface of each goat of three groups was inspected daily and decreases in M. ovinus were recorded. The rate of elimination in keds was assessed on the basis of decrease in keds count on the skin and hairs. The results revealed that complete absence of keds were observed in 6 and 7 days post-treatment with injection and pour-on routes, respectively. The results of present study showed that subcutaneous injection of ivermectin more rapidly eliminated M. ovinus than pour-on route. Both routes were 100% effective against this parasite in the goats. Ivermectin can be a drug of choice against M. ovinus in long-hair Iranian goats due to its high efficacy, easy applicability and wide safety margin.     

Comparison of phenotypic methods in predicting methicillin resistance in coagulase-negative Staphylococcus (CoNS) from animals

Phenotypic detection of methicillin resistance in coagulase-negative Staphylococcus (CoNS) of animal origin has been challenging due to the heterogeneous expression of mecA. To compare different phenotypic methods in predicting the mecA presence in CoNS, a total of 87 CoNS isolates from agricultural animals were analyzed in this study by agar dilution, disk diffusion, and broth microdilution. mecA was present in 81 CoNS isolates. Broth microdilution demonstrated the highest sensitivity of 100% in predicting the mecA presence, followed by 72.8% by agar dilution and 70.4% by disk diffusion. The results indicate that broth microdilution may be more suitable for predicting the presence of mecA in CoNS from animals than the other two methods, although staphylococcal species may also be a factor affecting the sensitivities of the methods as the top three staphylococcal species in this study were Staphylococcus lentus, Staphylococcus sciuri, and Staphylococcus xylosus (a total of 75 of 87).     

First report of thelaziosis (Thelazia anolabiata) in an Andean Cock of the Rock (Rupicola peruviana) from Peru

Thelazia anolabiata is a spirurid nematode living in the orbits of birds which can cause lacrimation, keratitis, conjunctivitis, and corneal ulcers. This species is reported for the first time from an Andean Cock of the Rock, Rupicola peruviana (Passeriformes: Cotingidae), from a zoo in Lima, Peru. Clinical signs of keratoconjunctivitis were resolved with the treatment of ivermectin, ciprofloxacin, and an epithelium regenerator, and the host is presently in good health. The nematodes were identified using the primary characteristics used to differentiate the species of this genus as lengths of spicules and other morphologic characteristics.     

Standardisation of a coproantigen reduction test (CRT) protocol for the diagnosis of resistance to triclabendazole in Fasciola hepatica

A sheep trial was performed to standardise a coproantigen reduction test (CRT) protocol for the diagnosis of resistance to triclabendazole (TCBZ) in Fasciola hepatica). The CRT employs the BIO K201 Fasciola coproantigen ELISA (Bio-X Diagnostics, Jemelle, Belgium) to test for the presence of F. hepatica coproantigens in a faecal sample. If it is coproantigen-positive, the CRT protocol recommends that faecal samples are re-tested for coproantigens at 14 days post-treatment (dpt), with negative testing at this point indicating TCBZ success. Initial work aimed to confirm the sensitivity of the BIO K201 ELISA for Fasciola infection and investigate whether coproantigens represent a robust reduction marker of TCBZ efficacy. Thirty-eight, indoor-reared sheep were artificially infected with F. hepatica isolates known to be susceptible (Cullompton) and resistant (Sligo) to TCBZ action, respectively. Treatment was administered at 12 weeks post-infection (wpi), with 2 sheep groups, infected with each isolate, culled at 2 and 4 weeks post-treatment (wpt), respectively. Necropsy was performed to confirm treatment efficacy. Individual faecal samples were collected twice-weekly throughout the trial period. Additional work focused on the effect of temperature on faecal sample collection and storage. Faecal samples collected from sheep positive for F. hepatica infection were sub-sampled and left at room temperature. Individual sub-samples were tested by ELISA on consecutive days and these readings compared to the original test result on the day of collection. In addition, ELISA values were compared between faecal sub-samples prepared on the day of sampling and post storage at -20°C. Also, an immunocytochemical study was performed to determine the tissue site of origin of the coproantigen protein in the fluke. Results showed that the BIO K201 ELISA was sensitive for Fasciola coproantigens, with coproantigens detectable from 5 wpi onwards. The suitability of coproantigens as a diagnostic marker of TCBZ efficacy was supported by the absence and presence of coproantigens in TCBZ-treated Cullompton (TCBZ-susceptible) and Sligo (TCBZ-resistant) F. hepatica infections at 2 and 4 wpt, respectively. Study results suggest that low to moderate temperature has little, if any, impact on coproantigen stability in faecal samples, but that higher temperatures may have. Immunolabelling for the coproantigen showed that it was specific to the gastrodermal cells of both adult and juvenile flukes.     

Pathogens of zoonotic and biological importance in roe deer (Capreolus capreolus): Seroprevalence in an agro-system population in France

Antibody prevalence for several infectious and parasitic diseases in a population of roe deer (Capreolus capreolus) inhabiting a mixed agricultural landscape (south of France) has been analyzed. Serological analyses with ELISA in 245 animals captured from 2008 to 2012 has been performed. We found a high prevalence of Toxoplasma gondii (46.4%), Chlamydophila abortus (17.27%) and Coxiella burnetii (11.26%) compared to other studies in Europe. Seroprevalence varied strongly among years for T. gondii (27–91%), C. abortus (0–42%) and C. burnetii (0–27%). T. gondii prevalence was lower in juvenile females, compared to juvenile males and adults of both sexes. Other pathogens had low prevalences: Neospora caninum (1.56%), Bovine herpesvirus 1 (1.17%, 2008/09; 1.1%, 2010/11), Mycoplasma agalactiae (1.45%, 2009/10), Mycobacterium avium subsp. paratuberculosis (0.9%) and Slow viruses (CAEV–MVV) (0.15%, 2008/10; 0%, 2011/12). Antibodies to bluetongue virus and pestiviruses were not found in any individual.     

Detection and molecular characterization of Theileria sp. in fallow deer (Dama dama) and ticks from an Italian natural preserve

The prevalence of piroplasms in a closed population of fallow deer (Dama dama L.) living in the Italian preserve of “Bosco della Mesola” - Ferrara (Mesola wood) was investigated. Blood samples and ticks were collected from 62 fallow deer. On microscopic observation, 28 (45.0%) blood samples were positive for piroplasms while PCR provided evidence for piroplasms infection in 47 (75.8%) fallow deer. The 67 ticks, collected from positive and negative animals, were identified as Ixodesricinus L., 1758 (89.6%) and Haemaphysalisconcinna Koch, 1844 (10.4%). At the PCR, four samples of I. ricinus were positive for piroplasms. The sequences of the 18S rRNA gene from both blood and ticks were identical and showed high identity (99.6%) with Theileria sp. 3185/02 (DQ866842) and Theileria capreoli (AY726011) from roe deer. Interestingly, the phylogenetical analyses evidenced differences between the Theileria strain from Mesola wood and the ones isolated in fallow deer from other Italian areas.     

Efficiency of entomopathogenic fungi in the control of eggs and larvae of the horn fly Haematobia irritans (Diptera: Muscidae)

The present study assessed the pathogenic effect of isolates E9, IBCB425 and IBCB159 of the Metarhizium anisopliae fungus, JAB06, JAB07 and AM09 of Beauveria bassiana, IBCB133 and CB75 of Isaria fumosorosea (=Paecilomyces fumosoroseus) and CG189 and CG195 of Isaria farinosa (=Paecilomyces farinosus) against eggs and larvae of the horn fly Haematobia irritans. Eggs were inoculated with suspensions containing 10⁶, 10⁷ and 10⁸ conidia ml⁻¹ of the fungal isolates and observed after 48 h to determine viability. In the larvae study, eggs were allowed to hatch into fresh bovine feces that had been treated with 10⁸, 10⁷ or 10⁶ conidia mg feces⁻¹. In both studies, 5 days after initial procedures, all formed pupae were transferred to an incubator at 27 ± 0.5 °C until the emergence of the adult flies was complete. The M. anisopliae isolates did not cause the death of H. irritans eggs, but they did promote the death of larvae that hatched from treated eggs, and therefore increased the total mortality. Isolate E9 promoted 100% mortality of treated larvae at a concentration of 10⁸ conidia ml⁻¹. For the B. bassiana isolates, no activity was observed against insect eggs or larvae. Both I. fumosorosea isolates promoted significant mortality (p < 0.05) of eggs at every concentration of conidia. Isolate CG195 of I. farinosa increased the mortality of larvae and pupae that hatched from treated eggs and promoted significant total mortality (p < 0.05) of the insect at every concentration of conidia.     

Identification of the tet(B) resistance gene in Streptococcus suis

The tetracycline resistance gene, tet(B), has been described previously in Gram negative bacteria. In this study tet(B) was detected in plasmid extracts from 17/111 (15%) Streptococcus suis isolates from diseased pigs, representing the first report of this resistance gene in Gram positive bacteria.     

Dynamics of shiga-toxin producing Escherichia coli (STEC) and their virulence factors in cattle

Starting at birth, twenty Holstein calves were housed individually, in groups of five and finally in one large freestall while fecal samples were collected weekly for 25 weeks. From each sample, twenty isolates of Escherichia coli were screened for 6 virulence markers including shiga-toxin 1, 2, intimin, enterohemolysin, the fimbrial antigen efa1 and the adhesin saa. Dynamic models of transmission of E. coli were used to model the transmission of different virulotypes between calves and the loss of the same virulotypes from the calves. It was found that, once E. coli encoding shiga-toxins in combination with enterohemolysin were transmitted and established in a calf, they tended to be eliminated less efficiently compared to E. coli without this combination of virulence markers. It was concluded that the presence of certain combinations of virulence markers coincided with persistence of E. coli in the bovine gastrointestinal tract. In addition, the combinations of stx with either eae or ehxA in E. coli have a greater impact on the loss rates than on the transmission rates.     

Phenotypic and genotypic characteristics of antimicrobial resistant Escherichia coli isolated from symbovine flies, cattle and sympatric insectivorous house martins from a farm in the Czech Republic (2006-2007)

The prevalence of antimicrobial resistant Escherichia coli was tested in symbovine flies and sympatric house martins (Delichon urbica) at a dairy farm. Antimicrobial resistant E. coli was detected in 89% (n = 147) of isolates from flies within a calf barn. Isolates with the same antimicrobial resistance phenotypes, genes, and pulsotypes were found between both fly and calf E. coli isolates, suggesting that the calves were the initial source of the antimicrobial resistant strains in fly isolates. Symbovine flies were considered as important reservoirs of antimicrobial resistant E. coli strains at a dairy farm, due to their intensive contact with cattle feces and manure. House martin fecal samples from the same farm contained 4.5% (n = 393) of antimicrobial resistant E. coli. House martin isolates displayed different macrorestriction profiles than fly isolates and the significance of house martins as a reservoir and vector of antimicrobial resistant E. coli appears low.     

Evaluation of the diagnostic accuracy of the modified agglutination test (MAT) and an indirect ELISA for the detection of serum antibodies against Toxoplasma gondii in sheep through Bayesian approaches

The diagnostic accuracies of the modified agglutination test (MAT) and indirect ELISA test for the detection of serum antibodies against Toxoplasma gondii in sheep were evaluated through Bayesian approaches on two populations of sheep created from three different groups of animals (T. gondii-aborted ewes, colostrums-deprived newborn lambs, and ewe-lambs and adult ewes with unknown T. gondii infection status). Tests showed a high degree of agreement (kappa statistic = 0.93; 95% confidence interval = 0.87, 0.98) and a significant specificity (Sp) correlation (gamma(Sp) = 0.26; 95% credibility interval = 0.017, 0.61). When prior information was used for all unknown parameters the posterior medians for the sensitivity (Se) and Sp of the MAT and ELISA were, respectively, 92.6% (95% credibility interval = 85.2, 96.9), 95.5% (89.9, 98.7), 90.5% (83.4, 95.6), and 97.8% (94.2, 99.5). These estimates remained similar when uninformative priors were included. The Se estimates of the MAT and ELISA were higher than those obtained on pigs in other study using the same approach (Se = 80.6% and Sp = 89.5% for the MAT, and Se = 71.5% and Sp = 85.5% for the ELISA [Georgiadis, M.P., Wesley, O.J., Gardner, I.A., Singh, R., 2003. Correlation-adjusted estimation of sensitivity and specificity of two diagnostic tests. Appl. Stat. 52, 63-78]. This finding supported the believe that test performances may vary when applied on different animal species. Thus, if these tests are planned to be used on animal species other than sheep or pigs, their diagnostic accuracy should be re-assessed to prevent biased inferences from their results.