DNA-based assessment of root lesion nematode infections in cereal roots.

Root lesion nematodes (RLN) of the genus Pratylenchus are causing significant damage in cereal production worldwide. Due to climate change and without efficient and environment-friendly treatments, the damages through RLNs are predicted to increase. Microscopic assessments of RLNs in the field and the greenhouses are time-consuming and laborious. As a result, cereal breeders have mostly ignored this pest. We present a method measuring RLN in infected cereal roots using a standardized PCR approach. Publicly available Pratylenchus neglectus primer combinations were evaluated. An optimal primer combination for RT-qPCR assay was identified to detect and quantify P. neglectus within infected cereal roots. Using the RT-qPCR detection assay, P. neglectus could be clearly distinguished from other plant parasitic nematodes. We could identify P. neglectus DNA in barley and wheat roots as low as 0.863 and 0.916 ng/µl of total DNA, respectively. A single P. neglectus individual was detected in water suspension and within barley and wheat roots. The RT-qPCR detection assay provides a robust and accurate alternative to microscopic nematode identification and quantification. It could be of interest for resistance breeding, where large populations must be screened to detect and quantify P. neglectus in farmer's fields.

Low-cost molecular methods to characterise gastrointestinal nematode co-infections of goats in Africa.

BACKGROUND: Veterinary diagnostics aid intervention strategies, track zoonoses, and direct selective breeding programs in livestock. In ruminants, gastrointestinal nematode (GIN) parasites are a major cause of production losses, but morphologically similar species limit our understanding of how specific GIN co-infections impact health in resource-limited settings. To estimate the presence and relative abundance of GINs and other helminths at the species level, we sought to develop a low-cost and low-resource molecular toolkit applied to goats from rural Malawi smallholdings. METHODS: Goats were subjected to health scoring and faecal sampling on smallholdings in Lilongwe district, Malawi. Infection intensities were estimated by faecal nematode egg counts with a faecal subsample desiccated for DNA analysis. Two DNA extraction methods were tested (low-resource magbead kit vs high-resource spin-column kit), with resulting DNA screened by endpoint polymerase chain reaction (PCR), semi-quantitative PCR, quantitative PCR (qPCR), high-resolution melt curve analysis (HRMC), and 'nemabiome' internal transcribed spacer 2 (ITS-2) amplicon sequencing. RESULTS: Both DNA isolation methods yielded comparable results despite poorer DNA purity and faecal contaminant carryover from the low-resource magbead method. GINs were detected in 100% of samples regardless of infection intensity. Co-infections with GINs and coccidia (Eimeria spp.) were present in most goats, with GIN populations dominated by Haemonchus contortus, Trichostrongylus colubriformis, Trichostrongylus axei, and Oesophagostomum columbianum. Both multiplex PCR and qPCR were highly predictive of GIN species proportions obtained using nemabiome amplicon sequencing; however, HRMC was less reliable than PCR in predicting the presence of particular species. CONCLUSIONS: These data represent the first 'nemabiome' sequencing of GINs from naturally infected smallholder goats in Africa and show the variable nature of GIN co-infections between individual animals. A similar level of granularity was detected by semi-quantitative PCR methods, which provided an accurate summary of species composition. Assessing GIN co-infections is therefore possible using cost-efficient low-resource DNA extraction and PCR approaches that can increase the capacity of molecular resources in areas where sequencing platforms are not available; and also open the door to affordable molecular GIN diagnostics. Given the diverse nature of infections in livestock and wildlife, these approaches have potential for disease surveillance in other areas.

[Temporal trends in disease burden of major human parasitic diseases in China from 1990 to 2019].

OBJECTIVE: To analysize the temporal trends in the disease burden of major human parasitic diseases in China from 1990 to 2019, so as to provide the evidence for improving the parasitic disease control strategy in China. METHODS: The disability-adjusted life years (DALYs) of malaria, intestinal nematode infections, schistosomiasis, food-borne trematodiases, cysticercosis and echinococcosis in China from 1990 to 2019 were captured from the Global Burden of Disease Study 2019 (GBD 2019), and age- and gender-specific DALYs of parasitic diseases were estimated. The temporal trends in DALYs of malaria, intestinal nematode infections, schistosomiasis, food-borne trematodiases, cysticercosis and echinococcosis were evaluated in China from 1990 to 2019 using average annual percent change (AAPC) with Joinpoint regression analysis. RESULTS: The DALYs were 643 836.42 person-years due to food-borne trematodiases, 156 853.03 person-years due to cysticercosis, 79 764.62 person-years due to schistosomiasis, 70 989.73 person-years due to intestinal nematode infections, 4 258.61 person-years due to echinococcosis and 264.86 person-years due to malaria in China in 2019, respectively. The overall DALYs of six parasitic diseases were higher among men (546 441.93 person-years) than among women (409 525.33 person-years), and were greater among adults at ages of 14 to 65 years (684 780.84 person-years) than among children at 14 years and lower (35 437.38 person-years) and the elderly at ages of 65 years and older (235 749.04 person-years). During the period from 1990 to 2019, food-borne trematodiases were the leading cause of DALYs among the six parasitic diseases, and cysticercosis shifted from the fourth leading cause in 1990 to the second leading cause of DALYs in China in 2019, while intestinal nematode infections shifted from the second leading cause in 1990 to the fourth leading cause of DALYs in 2019. The DALYs of major human parasitic diseases appeared an overall tendency towards a decline in China from 1990 to 2019, with the fastest drop seen in DALYs due to malaria (AAPC = -19.6%, P = 0.003), followed by due to intestinal nematode infections (AAPC = -8.2%, P < 0.001) and schistosomiasis (AAPC = -3.1%, P < 0.001), and a slow decline was seen in the DALYs of food-borne trematodiases (AAPC = -1.0%, P < 0.001), while there were no significant decrease in the DALYs of echinococcosis (AAPC = -0.5%, P = 0.264) and the DALYs of cysticercosis appeared a tendency towards a rise (AAPC = 0.7%, P < 0.001). CONCLUSIONS: The disease burden of major human parasitic diseases appeared an overall tendency towards a decline in China from 1990 to 2019, with a high disease burden seen due to food-borne parasitic diseases, no remarkable reduction seen in echinococcosis, and a tendency towards a rise seen in cysticercosis. It is recommended to focus on echinococcosis control, and continue to consolidate the control achievements of other major human parasitic diseases in China; meanwhile, the surveillance and prevention of food-borne parasitic diseases should be reinforced.

Economic impact of gastrointestinal nematodes in Morada Nova sheep in Brazil.

This study evaluated the economic impact of gastrointestinal nematode (GIN) infection in Morada Nova lambs under different parasite chemical control conditions. For this, 246 lambs, in the rainy and dry season, were randomized into groups according to their anthelmintic treatment with levamisole: control (CT: no treatment); routine treatment (RT: treated every 42 days); and targeted selective treatment (TST: treated according to the average daily weight gain, DWG). From 63 days of age (D63) to D210, the lambs were weighed and monitored for GIN infection parameters. Spending on anthelmintics in the production system was 1.3% of the total economic result. The economic result per animal (R$ 5.00 = US$ 1.00) was higher in the RT group, amounting to US$ 6.60 in the rainy and US$ 5.69 in the dry season, due to higher DWG. Thus, RT presented economic results 14.4% and 10.9% higher than CT, and 7.2% and 1.9% higher than TST, in the rainy and dry season, respectively. However, fast development of resistance made RT unfeasible. Here, the economic impact of GIN infection on a national scale is discussed, demonstrating its importance and the impossibility of profitable and sustainable sheep production without adequate control.

Preliminary Assessment of Body Condition Score as a Possible Marker for the Targeted Selective Treatment of Dairy Sheep Against Gastrointestinal Nematodes.

PURPOSE: In the fight against anthelmintic resistance, targeted selective treatments (TSTs), where only a small percentage of a flock receives treatment, have become increasingly popular. Overall, implementation of such treatments can be based on various parameters including Body condition score (BCS). As infection with non-bloodsucking nematodes, frequently encountered on sheep farms in the central Mediterranean basin, commonly causes bodyweight reduction, the aim of this research is to evaluate the effectiveness of BCS as a parameter for the implementation of TSTs in lactating dairy sheep with subclinical gastrointestinal nematode (GIN) infections from the island of Sardinia, Italy. METHODS: Faecal samples from 1012 ewes divided into 2 groups (third and fifth month of lactation) were collected and their BCS recorded. Faecal egg counts and coprocultures were performed for the assessment of the GIN burden and identification of present species. RESULTS: An overall GIN prevalence of 85.4% with a mean eggs per gram (EPG) of faeces of 210.1 ± 347.3 was found. Teladorsagia spp. and Trichostrongylus spp. were the GIN genera most identified. Overall, animals with the lowest BCS had the highest EPG values and a negative correlation (r = - 0.163) between the EPG values and BCS of the studied animals was found, which was most significant for older sheep. CONCLUSION: This research confirmed BCSs and EPG values for GIN in sheep to be negatively correlated, particularly in older ewes. Application of TSTs for lactating sheep with a BCS < 2.25, especially to older ewes, could be beneficial in case of subclinical GIN infections, although further studies are needed to work out precise recommendation.

Economic impact of gastrointestinal nematodes in Morada Nova sheep in Brazil / Impacto econômico de nematódeos gastrintestinais em ovinos Morada Nova no Brasil

Abstract This study evaluated the economic impact of gastrointestinal nematode (GIN) infection in Morada Nova lambs under different parasite chemical control conditions. For this, 246 lambs, in the rainy and dry season, were randomized into groups according to their anthelmintic treatment with levamisole: control (CT: no treatment); routine treatment (RT: treated every 42 days); and targeted selective treatment (TST: treated according to the average daily weight gain, DWG). From 63 days of age (D63) to D210, the lambs were weighed and monitored for GIN infection parameters. Spending on anthelmintics in the production system was 1.3% of the total economic result. The economic result per animal (R$ 5.00 = US$ 1.00) was higher in the RT group, amounting to US$ 6.60 in the rainy and US$ 5.69 in the dry season, due to higher DWG. Thus, RT presented economic results 14.4% and 10.9% higher than CT, and 7.2% and 1.9% higher than TST, in the rainy and dry season, respectively. However, fast development of resistance made RT unfeasible. Here, the economic impact of GIN infection on a national scale is discussed, demonstrating its importance and the impossibility of profitable and sustainable sheep production without adequate control.

Resumo Este estudo avaliou o impacto econômico da infecção por nematoides gastrintestinais (NGI), em cordeiros Morada Nova, sob diferentes condições de controle químico dos parasitas. Para isso, 246 cordeiros, na estação chuvosa e seca, foram randomizados em grupos de acordo com o tratamento com levamisol: controle (TC: sem tratamento); tratamento rotineiro (TR: tratado a cada 42 dias); e tratamento seletivo direcionado (TST: tratado de acordo com o ganho de peso médio diário, GMD). Dos 63 dias de idade (D63) ao D210, os cordeiros foram pesados ​​e monitorados quanto aos parâmetros de infecção por NGI. O gasto com anti-helmínticos no sistema produtivo foi de 1,3% do resultado econômico total. O resultado econômico por animal (R$ 5,00 = US$ 1,00) foi maior no grupo RT, totalizando US$ 6,60 na estação chuvosa e US$ 5,69 na seca, devido ao maior GMD. Assim, o RT apresentou resultados econômicos 14,4% e 10,9% superiores ao TC, e 7,2% e 1,9% superiores ao TST, no período chuvoso e seco, respectivamente. Entretanto o rápido desenvolvimento de resistência inviabiliza o TR. O impacto econômico da infecção por NGI em escala nacional são aqui discutidos, demonstrando sua importância e a impossibilidade de uma ovinocultura lucrativa e sustentável sem o controle adequado.

[Prevalence of human soil-borne nematode infections in Yunnan Province: a cross-sectional study in 2015].

OBJECTIVE: To investigate the current prevalence of human soil-borne nematode infections in Yunnan province, so as to provide the scientific evidence for formulating the soil-borne nematodiasis control strategy in the province. METHODS: In 2015, a total of 20 survey sites were sampled in 10 counties (cities) of Yunnan Province using the stratified cluster random sampling method. Stool samples were collected from all local permanent residents at ages of one year and older in each survey site, and the soil-borne nematode eggs were identified using the modified Kato-Katz technique and the egg number was counted. In addition, the hookworm species was identified using the filter-paperculture method, and Enterobius vermicularis eggs were detected using the cellophane tape method in children at ages of 3 to 6 years. RESULTS: A total of 5 067 residents received stool examinations, and 950 residents were detected with soil-borne nematode infections, with an overall prevalence rate of 18.75%. The prevalence of Ascaris lumbricoides, Trichuris trichura and hookworm was 7.52%, 8.47% and 9.02%, respectively. Among 446 children detected using the cellophane tape method, 5 children were detected with E. vermicularis infections. Among the 160 residents with hookworm infections, there were 139 residents with Necator americanus infections (86.88%), 16 with A. duodenale infections (10.00%) and 5 with mixed infections (3.12%). Mild A. lumbricoides (67.98%, 259/381), T. trichura (88.58%, 380/429) and hookworm infections (94.53%, 432/457) were predominant. Among the four ecological zones, the highest prevalence of human soilborne nematode infections was found in the East Tibet-South Sichuan Ecological Zone (31.79%), and among the 10 survey counties (cities), the greatest prevalence was seen in Gongshan Derung and Nu Autonomous County (50.13%), while the lowest prevalence was found in Ninglang Yi Autonomous County (0.40%). The prevalence of human soil-borne nematode infections was 5.67% (43/759), 26.67% (610/2 287) and 14.70% (297/2 021) in high-, moderate- and low-economic-level regions, respectively. There were no significant differences in the prevalence of human soil-borne nematode infections in terms of ecological regions, survey counties (cities) or economic development levels (χ2 = 342.20, 814.60 and 201.34, all P < 0.05). There was no significantdifference in the prevalence of human soil-borne nematode infections between male (18.21%, 441/2 422) and female residents (19.24%, 509/2 645) (χ2 = 0.89, P > 0.05), and soil-borne nematode infections were detected in residents at all age groups, with the greatest prevalence found in residents at ages of 1 to 9 years (25.88%). In addition, the highest prevalence of soil-borne nematode infections was seen in residents with the Dulong Ethnic Minority (82.09%), in preschool children (25.06%) and in illiterate residents (24.80%), and there was no age-, ethnicity-, occupation- or education level-specific prevalence of soil-borne nematode infections detected (χ2 = 46.50, 1 016.96, 36.33 and 52.43, all P < 0.05). CONCLUSIONS: The prevalence of human soil-borne nematode infections remains high in Yunnan Province. The management of soil-borne nematodiasis requires to be reinforced among low-age children, farmers, old people and residents with low educations levels or ethnic groups.

The identification and semi-quantitative assessment of gastrointestinal nematodes in faecal samples using multiplex real-time PCR assays.

BACKGROUND: The diagnosis of gastrointestinal nematode (GIN) infections in ruminants is routinely based on morphological/morphometric analysis of parasite specimens recovered by coprological methods, followed by larval culture (LC) techniques. Such an approach is laborious, time-consuming, requires a skilled expert, and moreover suffers from certain limitations. Molecular tools are able to overcome the majority of these issues, providing accurate identification of nematode species and, therefore, may be valuable in sustainable parasite control strategies. METHODS: Two multiplex real-time polymerase chain reaction (PCR) assays for specific detection of five main and one invasive GIN species, including an internal amplification control to avoid false-negative results, were designed targeting SSU rRNA and COI genetic markers, as well as established ITS1/2 sequences. The assays were optimized for analysis of DNA extracted directly from sheep faeces and verified for Haemonchus contortus, Teladorsagia circumcincta, Trichostrongylus colubriformis, Nematodirus battus, Chabertia ovina, and Ashworthius sidemi. Semi-quantitative evaluation of infection intensity was enabled using a plasmid construct and a dilution series of sheep faeces with a known number of nematode eggs. Assays were tested on 44 individually collected faecal samples from three farms, and results were compared to those from faecal egg counts (FEC) using the concentration McMaster technique and LC. RESULTS: Multiplex real-time PCR assays showed great specificity to target nematodes. During the analysis of faecal samples, the assays proved to have higher sensitivity in strongylid-type egg detection over FEC by revealing three false-negative samples, while showing moderate agreement in evaluation of infection intensity. The multiplex assays further clarified GIN species identification compared to LC, which had confused determination of Teladorsagia spp. for Trichostrongylus spp. CONCLUSIONS: Our multiplex assays proved to be a rapid and accurate approach enabling simultaneous and reliable GIN species identification from faeces and semi-quantitative estimation of the number of eggs present. This approach increases diagnostic value and may add a high degree of precision to evaluation of anthelmintic efficacy, where it is important to identify species surviving after treatment.

Assessment of in vitro anthelmintic activity and bio-guided chemical analysis of BRS Boyrá pineapple leaf extracts.

Species of the Bromeliaceae are known for their pharmacological actions, including anthelmintic effects. The aim of this study was to investigate the in vitro anthelmintic activity of extracts and fractions of BRS Boyrá pineapple leaf against the eggs and infective larvae of gastrointestinal nematodes (Trichostrongylidae) of goats and to identify the compounds involved in this activity. Crude methanol, hexane, dichloromethane, ethyl acetate and residual hydromethanol extracts were investigated by quantitative analysis of phenolic and flavonoid contents, antioxidant activity, anthelmintic activity against gastrointestinal nematodes of goats. The extracts were submitted to chromatographic methods for substance isolation and spectrometric techniques to identify their structures. The anthelmintic activity was performed by in vitro assays with eggs and larvae of nematodes obtained from naturally infected donor goats. All extracts contained phenolic (2.22-14.12 g of gallic acid equivalent per 100 g of dry extract) and flavonoid compounds (0.13-1.45 g of quercetin equivalent per 100 g of dry extract). Bio-guided fractionation of the BRS Boyrá pineapple leaves showed high antioxidant activity (EC50 for DPPH of 2.16-21.38 mg mL-1 and inhibition of co-oxidation of ß-carotene of 36.40-74.86%) and anthelmintic activity (15.69-100% inhibition of egg hatching). The ethyl acetate extract exhibited greatest activity in all assays. Through chromatographic column analysis it was possible to isolate three substances: ß-sitosterol and stigmasterol mixture in dichloromethane and hexane extracts, identified by NMR and p-coumaric acid in the ethyl acetate extract, identified by HPLC-DAD. The isolated p-coumaric acid exhibited high ovicidal effect against goat gastrointestinal nematodes (IC50: 0.12 mg mL-1) and can be considered the active substance of the ethyl acetate extract. This study revealed for the first time that the pineapple BRS Boyrá possesses inhibitory activity against gastrointestinal nematodes (Haemonchus spp., Oesophagostomum spp. and Trichostrongylus spp.), and that p-coumaric acid is an important bioactive.

Effects of long-term indiscriminate use of macrocyclic lactones in cattle: Parasite resistance, clinical helminthosis, and production losses.

Our aims were to describe a case of clinical helminthosis caused by parasite resistance to macrocyclic lactones (MLs) after the long-term frequent use of these drugs in a cattle herd, and to evaluate the production losses prevented by the use of an effective anthelmintic treatment to control these resistant gastrointestinal nematodes (GINs). A case of clinical helminthosis culminating in the death of steers was investigated, the history of the antiparasitic treatments used during an 11-year period in the herd was assessed, and an efficacy test involving seven different drugs was performed. Thereafter, two groups of heifers naturally infected by ML-resistant GINs were formed and strategically treated with either a highly effective (levamisole) or less effective drug (doramectin) over a 9-month period. The heifers were evaluated monthly based on eggs per gram of feces (EPG) counts and liveweights. An evaluation of the history of parasite control in the farm revealed that MLs were used in 96.5% of the treatments aimed at controlling GINs, ticks, and myiasis in the herd. The efficacy test showed the presence of GINs resistance to all the MLs tested. However, levamisole and albendazole sulphoxide were highly effective against these parasites. Heifers treated with levamisole gained 12.1 kg more liveweight on average, compared to those treated with doramectin. Thus, we conclude that indiscriminate and long-term use of MLs in the studied herd led to the failure of GINs control, a critical situation resulting in significant production losses, and a surge of clinical helminthosis in young cattle. In addition, we showed increase in liveweight gain due to using a highly effective drug, in comparison to an ML, during a 9-month period, in heifers naturally infected by ML-resistant GINs.

Assessment of anthelmintic efficacy against cattle gastrointestinal nematodes in western France and southern Italy.

Our objective was to measure the efficacy of ivermectin (IVM) and benzimidazoles (BZ, i.e. fenbendazole and albendazole) in 15 cattle farms in western France and southern Italy. A total of 11 groups were treated with IVM and 11 with BZ. Efficacy was assessed by calculating the percentage of faecal egg count reduction (%FECR) using the pre- and post-treatment arithmetic means. Anthelmintic resistance was considered to be present when the %FECR was <95% and the lower limit of the 95% confidence interval <90%. For IVM, the percentages of FECR ranged from 73% to 100%. Lack of efficacy to IVM was detected in two farms out of four in France, but was not detected in any of the seven farms in Italy. For BZ, the percentages of FECR ranged from 95% to 100%. No case of BZ resistance was detected in the five farms in France and the six farms in Italy.

Biology, Epidemiology, Diagnosis, and Management of Anthelmintic Resistance in Gastrointestinal Nematodes of Livestock.

Control of gastrointestinal nematodes has been based on anthelmintics. However, this strategy is unsustainable owing to anthelmintic resistance. Parasitic nematodes have biologic and genetic features that favor the development of drug resistance, making the emergence of resistant nematodes inevitable. The rate of resistance development is affected controllable factors. There is a need to change the paradigm of how gastrointestinal nematodes are controlled to decrease the rate at which resistance develops. This article reviews the biology and prevalence of anthelmintic resistance, and provides recommendations for diagnosing resistance and for strategies that should be implemented to reduce the development of resistance.

Assessment of oxidative/nitrosative stress biomarkers and DNA damage in Teladorsagia circumcincta following exposure to zinc oxide nanoparticles.

Drug resistance to helminth parasites is one of the most serious problems to threaten the livestock industry. The problem also poses a major threat to public health. Therefore, novel and safe agents should urgently be investigated to control parasitic infections. The current study was conducted to evaluate the possible antiparasitic effects of zinc oxide nanoparticles (ZnO-NPs) on one of the most prevalent gastrointestinal nematodes, Teladorsagia circumcincta. The worms were incubated with various concentrations of ZnO-NPs: 1, 4, 8, 12 and 16 ppm for 24 hours. Mobility and mortality of the parasites were recorded at four-hour intervals. At the endpoint, several biomarkers of oxidative/nitrosative stress, including superoxide dismutase, glutathione peroxidase and catalase, as well as lipid peroxidation, protein carbonylation, total antioxidant status, nitric oxide contents and DNA damage, were measured in the homogenized samples. ZnO-NPs showed significant anthelminthic effects, depending on time and concentration. Furthermore, the nanoparticle induced severe oxidative/nitrosative stress and DNA damage. ZnO-NPs could be considered as a novel and potent anthelminthic agent.

Assessment of targeted selective treatment criteria to control subclinical gastrointestinal nematode infections on sheep farms.

Control of sheep gastrointestinal nematode (GIN) infections mostly relies on the use of anthelmintics. Refugia-based control strategies as targeted selective treatments (TST) can delay anthelmintic resistance development, but the optimal decision criteria for selecting individuals to be treated in subclinical infection scenarios remain unclear. The present study aimed to evaluate the suitability of body condition score (BCS) and faecal egg counts (FEC) as treatment indicators and to determine their optimized threshold values for treatment in TST by determining the relationships of BCS, FEC and anthelmintic treatment with several productive parameters in pre-mating and pre-partum periods at an individual level. Deworming in pre-mating period increased BCS gain, but its magnitude was directly associated with strongyle FEC before treatment. Deworming also increased fertility in ewes with BCS < 2.75 and the proportion of lambing ewes that got pregnant during the first ovulation cycle. Nevertheless, treatment did not yield productive benefits in ewes with higher BCS values. On the other hand, deworming in the pre-partum lowered lamb peri-partum mortality and increased lamb weight at birth and the growth of lambs during lactation, especially in lambs born from ewes with strongyle FEC > 400 epg before treatment. These results showed that FEC and especially BCS can be potential decision criteria for the implementation of TST in these types of scenarios. The TST scheme derived from the present results on the control of GIN infections should include anthelmintic treatment of ewes with BCS lower than ≈ 3 approximately five weeks before mating and lambing. In the pre-lambing period, treatment may be unnecessary if flock mean strongyle FEC is lower than ≈ 200 epg, as the proportion of ewes with individual FEC > 400 epg would be very low. The results suggest that implementation of this TST scheme would provide benefits, such as the improvement of productivity, a rational management of parasites in refugia, and preservation of future efficacy of anthelmintics, in comparison to traditional deworming schemes.

Economic losses caused by the use of low-efficacy anthelmintic drugs in growing heifers.

The aim of this study was to compare the economic revenue related to the use of low- or high-efficacy anthelmintic drugs within suppressive or strategic schemes of treatment in growing heifers. Heifers raised in a semi-intensive grazing system in southern Brazil were used. Levamisole and ivermectin were selected as the high- and the low-efficacy drugs, respectively, based on a previous efficacy test. Subsequently, these drugs were used within strategic (Strat; four times per year) or suppressive (Supp; once a month) treatment regimens in the heifers, and their liveweight and eggs per gram of feces counts were monthly evaluated during a 13-month period. The total costs of the treatments and their cost-benefit ratio in regard to liveweight gain were calculated. Final mean liveweight gains (kg) observed were 126.7 (Strat-Low), 133.6 (Supp-Low), 141.3 (Strat-High), 142.9 (Supp-High), and 125.8 (Control). Treatments with a high-efficacy drug resulted in monetary gains of US$ 19.56 (Strat-High) and US$ 14.98 (Supp-High), but Supp-Low and Strat-Low treatments caused economic losses. Total cost of the efficacy test (US$ 374.79) could be paid by the additional liveweight gain of 20 heifers from the Strat-High group. These results showed that it would be preferable not to treat the heifers against GIN if compared with treating them with a low-efficacy drug. In addition, we showed that the use of four treatments per year with a high-efficacy drug-selected by efficacy test-resulted in a profitable management to control GIN in growing heifers raised in a semi-intensive gazing system in southern Brazil.

Looking for insight: The role of parasite tester opinions in farm management.

Gastrointestinal nematode (GIN) infections cause millions of dollars of economic loss annually. Increasing cases of anthelmintic resistance have resulted in calls for restricted drug use and implementation of sustainable management practices to slow the rate of resistance. The limited uptake of available management systems and advice has sparked multiple surveys into the psychology and behaviours preventing uptake. These surveys have looked mainly at the farmers, the majority of whom have reported they rely on the advice of their local veterinarian or suppliers for treatment and management. However, there is little research into the psychology of veterinarians and people performing animal health testing on this topic. In the current study, a short survey of people performing faecal egg counts on animals was conducted. The survey focused on identifying areas for improving diagnosis to encourage uptake, and found other areas of interest worth further investigation. Respondents most frequently named manual labour as the main contributor to the cost of testing (65% of respondents) with analysis (42%) and sample preparation (32%) being the main contributors to time. In the survey comments, there was little consistency or commonality in the issues raised. The disparity between onsite and laboratory testers is an area worth investigation, particularly into how to co-ordinate behaviour and advice between proactive farmers and parasitology/veterinary services. Further investigation could provide better insight into how to encourage and maintain sustainable practices on farms.

Rapid assessment of faecal egg count and faecal egg count reduction through composite sampling in cattle.

BACKGROUND: Faecal egg counts (FEC) and the FEC reduction test (FECRT) for assessing gastrointestinal nematode (GIN) infection and efficacy of anthelmintics are rarely carried out on ruminant farms because of the cost of individual analyses. The use of pooled faecal samples is a promising method to reduce time and costs, but few studies are available for cattle, especially on the evaluation of different pool sizes and FECRT application. METHODS: A study was conducted to assess FEC strategies based on pooled faecal samples using different pool sizes and to evaluate the pen-side use of a portable FEC-kit for the assessment of FEC on cattle farms. A total of 19 farms representing 29 groups of cattle were investigated in Italy and France. On each farm, individual faecal samples from heifers were collected before (D0) and two weeks after (D14) anthelmintic treatment with ivermectin or benzimidazoles. FEC were determined individually and as pooled samples using the Mini-FLOTAC technique. Four different pool sizes were used: 5 individual samples, 10 individual samples, global and global on-farm. Correlations and agreements between individual and pooled results were estimated with Spearman's correlation coefficient and Lin's concordance correlation coefficients, respectively. RESULTS: High correlation and agreement coefficients were found between the mean of individual FEC and the mean of FEC of the different pool sizes when considering all FEC obtained at D0 and D14. However, these parameters were lower for FECR calculation due to a poorer estimate of FEC at D14 from the faecal pools. When using FEC from pooled samples only at D0, higher correlation and agreement coefficients were found between FECR data, the better results being obtained with pools of 5 samples. Interestingly, FEC obtained on pooled samples by the portable FEC-kit on-farm showed high correlation and agreement with FEC obtained on individual samples in the laboratory. This field approach has to be validated on a larger scale to assess its feasibility and reliability. CONCLUSIONS: The present study highlights that the pooling strategy and the use of portable FEC-kits on-farm are rapid and cost-effective procedures for the assessment of GIN egg excretion and can be used cautiously for FECR calculation following the administration of anthelmintics in cattle.

Comparison of the efficacy of crude methanolic extracts of Cassia occidentalis and Euphorbia hirta with levamisole-HCL against gastrointestinal nematodes of economic importance to goat production in Uganda.

Natural compounds from medicinal plants provide safe and sustainable alternatives to synthetic anthelmintics. In this study, we assessed in vitro and in vivo anthelmintic activity of Cassia occidentalis (NH-A) and Euphorbia hirta (NH-B) and compared it with levamisole-HCl. The shoots of NH-A and whole plant of NH-B were used to prepare extracts using 70% methanol which were used in the in vitro and in vivo assays. In vitro assays of crude methanolic extracts (CMEs) of NH-A and NH-B on larvae of mixed gastrointestinal nematodes (GINs) of goats revealed mortalities of 95.7% (at 100 mg ml-1) and 98.1% (at 50 mgml-1) 24 h postexposure. In vivo assays of NH-A administered orally at doses of 100, 300, 900, and 2700 mg kg-1 bwt revealed dose- and time-dependent anthelmintic effects in goats experimentally infected with mixed species of GINs. NH-B exhibited similar properties when administered at doses of 50, 100, 200, and 400 mg kg-1 bwt. Both NH-A (900 mg kg-1 bwt) and levamisole (7.5 mg kg-1 bwt) achieved a 100% reduction in fecal egg count per gram (EPG) on day 21 and day 14 respectively posttreatment. NH-B (400 mg kg-1 bwt) achieved 93.1% and 86.1% reduction in fecal EPG 7 and 14 days postexposure respectively compared with 88.2% and 82.3% reduction with levamisole-HCl 7 and 14 days postexposure. Our results show that extracts of both plants can disrupt helminth lifecycles by suppressing the egg-laying capacity in adult worms but also kill their infective larvae. Future studies should aim at establishing synergies or antagonisms between the two plant extracts and further development for control of helminths in goats.

Prevalence, economic assessment, and risk factors of gastrointestinal nematodes infecting herds in tropical, dry and temperate climate regions in Mexico.

This study investigated exposure to gastrointestinal (GI) nematodes on dairy cattle farms by antibody level determination in bulk tank milk (BTM) samples and its influence on production to detect the risk factors for infection in different climate regions in three states of Mexico. From January to April 2017, BTM samples were collected from 1058 dairy cattle herds and used to establish three Köppen climate classes (tropical, dry and temperate) and states of Mexico. A questionnaire on farm management was applied. The overall herd prevalence of parasites was 67.20%. The highest percentage of positive herds was detected in Veracruz state (78.45%). In addition, the highest prevalence among the climate regions was found in the tropical climate (78.59%). In general, production losses were approximately 1.37-1.78 kg of milk/cow per day. The annual costs of milk production losses per farm were estimated for three different climate regions, ranging between $5541.49 and 6982.50 US$, and those in the three states varied between 5974.10 and 8660.06 US$. The costs for anthelmintic treatments for young stock and adult cows ranged between 57.51 and 192.75 US$, respectively, among the three climate regions and between 46.02 and 189.49 US$, respectively, among the three states. The overall annual costs of milk yield loss per cow were estimated to be 150.74 US$ for the climate regions and 190.54 US$ for the three states of Mexico, followed by the treatment costs for young stock (4.02 US$) and adult cows (3.99 US$). The results suggest that the economic losses due to GI nematodes in Mexican dairy herds are approximately 248 million US$ per annum. Four final models were built based on multivariate logistic regression for potential statistical association from the ELISA results using climatic/environmental and management factors so that each model used different risk factors that were significantly associated with helminth infections in dairy herds.

Impact of gastrointestinal parasitism on dry matter intake and live weight gain of lambs: A meta-analysis to estimate the metabolic cost of gastrointestinal nematodes.

This study aimed to estimate the effect of gastrointestinal nematodes (GIN) on the productive performance assessed by the live-weight change (LWC) of lambs, and the metabolic cost associated with parasitism by means of a meta-analysis. Data used in the meta-analyses were obtained from twenty papers selected using the following criteria: (a) lambs with (I) and without (NI) GIN; (b) lambs fed ad libitum; (c) LWC data; (d) feed consumption data; and (e) chemical composition of diets. The effect of diet composition (crude protein [CP] and metabolizable energy [ME]) on dry matter intake (DMI), and the effect of composition and nutrient intake (DMI and CP intake [CPI]) and ME intake (MEI) on LWC was evaluated using respective regression analyses. The metabolic cost of worm burden was determined as the difference in CP and ME requirements between NI and I lambs for each adult parasite. The CP and ME cost were evaluated for lambs at two different levels: maintenance level 0 g LWC and 100 g LWC. The worm burden had an impact on the DMI and LWC of lambs (P < 0.05). The association of CP x infection level was the best predictor of DMI. The association between MEI and infection level were the best predictors of LWC. The metabolic cost of GIN increased to 0.30 mg CP/kg LW0.75 and 0.0056 kJ ME/kg LW0.75 for each adult parasite. The metabolic cost was not modified by diet quality (maintenance level 0 g LW/day or 100 g LW/day). In conclusion, GIN has a negative effect on DMI and LWC. The metabolic cost of GIN infection can be covered by supplying the additional requirements for protein and energy in the diet of infected lambs.