Anthelmintic Activity of Tanacetum vulgare L. (Leaf and Flower) Extracts against Trichostrongylidae Nematodes in Sheep In Vitro.

Due to the high prevalence of gastrointestinal nematodes in sheep, the growing anthelmintic resistance, and the development of organic farming systems, sustainable alternatives are being sought. One such method is phytotherapy. This study aimed to evaluate the in vitro ovicidal and larvicidal activity of extracts of tansy (Tanacetum vulgare L.) growing in Latvia on gastrointestinal nematodes (Trichostrongylidae) in sheep. The leaves and flowers of the tansy were extracted separately in 70%, 50%, and 30% ethanol and acetone. Six concentrations were prepared from each extract 500 mg/mL, 200 mg/mL, 100 mg/mL, 50 mg/mL, 20 mg/mL, and 10 mg/mL. In vitro egg hatching test and micro-agar larval development test were performed. Extracts of tansy have strong larvicidal activity. The highest percentage of larvae inhibition for most of the extracts was 100%, but for egg inhibition, it was 95.8% for the 200 mg/mL concentration of 50% acetone and 93.3% for the 500 mg/mL concentration of 50% ethanol leaf extracts. All tansy extracts had ovicidal and larvicidal activity against Trichostrongylidae in sheep.

Prevalence of susceptibility to Cryptosporidium spp. among dairy calves with different feeding regimens with an emphasis on the feeding of transition milk.

Background and Aim: Colostrum composition and importance for newborn organisms were repeatedly studied. However, the interest in transitional milk usefulness is weak and recommendations concerning transition milk intake are not developed. The aim of this study was to evaluate whether transition milk intake after colostrum consumption affects the chances of calf infection with Cryptosporidium spp. Materials and Methods: We collected data for Cryptosporidium spp. infection from calves (n=425) divided into three groups: The first group - supervised colostrum and transition milk intake; the second group - supervised colostrum and whole milk intake; and the third group - not supervised colostrum and whole milk intake. To detect oocysts of Cryptosporidium spp. in feces, the flotation method was used, and slides were stained using the modified Ziehl-Neelsen method. Generalized linear mixed modeling was conducted to determine whether the explanatory variable - the management of colostrum and transition milk feeding with three categories (three research groups) - was related to the probability of calves incurring infection with Cryptosporidium spp. Results: In the first group, 26.1% of calves were positive for the presence of Cryptosporidium spp. oocysts, in the second - 37.2%, and in the third - 44.1%. Statistical data analysis showed that calves who did not receive transition milk after colostrum consumption had increased chances of having Cryptosporidium spp. (by 1.90-2.47 times on average). The main results showed that the management of colostrum and transition milk feeding is related to Cryptosporidium spp. infection, indicating that both colostrum and transitional milk play a significant role in controlling pathogenic infections. Conclusion: The most effective management of colostrum and transition milk feeding against Cryptosporidium spp. infection is the timely intake of an adequate amount of colostrum followed by transitional milk consumption for at least 2 weeks before weaning from the dam.

Parasitofauna and current status of anthelmintic resistance in Latvian sheep farms.

Background and Aim: Parasitic invasions, especially gastrointestinal nematodes, are widespread and are one of the main problems in sheep farms. For this reason, sheep are dewormed more often than other livestock species, often several times a year. Concerns about antiparasitic resistance from the farmers and veterinarians have arisen because, on some farms, antiparasitic drugs are used routinely for very long periods. There are no data available on anthelmintic resistance in gastrointestinal nematodes in sheep in Latvia. Our work aimed to determine the most common endoparasites in sheep and the degree of anthelmintic resistance on sheep farms in Latvia. Materials and Methods: All sheep (577) underwent a coprological examination before the start of the study, and only sheep diagnosed with more than 200 McMaster eggs per gram of feces were included in the study. A fecal egg count reduction (FECR) test was performed on 20 sheep flocks in Latvia. Results: In Latvia, sheep were most commonly infected with Eimeria spp. 97% (confidence interval [CI] 95% 96-98). The second most commonly diagnosed species were Trichostrongylidae 91% (CI 95% 89-93) and Strongyloides spp. 76% (CI 95%, 72-79). The ivermectin (IVM) FECR was 0.74% (0.73-0.74), showing resistance in all sheep farms included in this study. Albendazole (ABZ) FECR 0.89% (0.88-0.89) was effective. Conclusion: This study showed that the most popular deworming drugs (ivermectin, albendazole) in Latvia are ineffective in sheep. Additional studies on the use of IVM+ABZ combinations for deworming sheep should be performed.

Most common inappropriate drug usage factors in anthelmintic treatment on sheep farms in Latvia.

Background and Aim: There is little understanding about antiparasitic drug prescription trends and implementation to reduce possible drug overuse or misuse worldwide. This study aimed to review sheep parasite control strategies and antiparasitic drug use habits in Latvia. To the best of the author's knowledge, this is the first study in the world that describes how antiparasitic drugs are used and what are the most common drug usage errors in a sheep farm. Materials and Methods: A semi-structured questionnaire was designed to collect relevant information from face-to-face interviews to assess 22 sheep farmers' knowledge and management procedures in farms. We collected information about animal feeding, herding, parasite diagnostics, and antiparasitic drug usage. The questionnaire summary included information on pasture use, parasite control management, and anthelmintic drug choice/use. Results: Only 36% of farms regularly managed parasite control by analyzing fecal samples for parasites, but prophylactic dewormingwas employed in all farms. Ivermectin, albendazole, levamisole, and monepantel were used on the farms and most of the farms were multidrug users; 77.3% of the farms used albendazole and 72.7% used ivermectin. Conclusion: The results indicated a lack of parasitological examination and parasite control of the flock, mostly empiric drug selection, incorrect dosing, inaccurate drug administration, drug storage, and use errors. A proactive approach to herd health planning, regular parasitic control, and prophylactic measures may benefit farmers and veterinarians.

Parity of Calving Influences the Likelihood of Calves Having Cryptosporidium spp.

The effect of colostrum on calves' health status was intensively studied, while the role of transition milk was left underestimated. The common practice is to feed calves with an adequate amount of colostrum immediately after calving and soon after feeding calves are weaned from dams. In this research, calves were not weaned from dams for at least 2 weeks receiving both colostrum and transition milk on demand. Thus, we have recreated natural feeding conditions for calves' development. We used a stratified sample method to test whether the size of the dairy cattle farms, breed, parity number, season of calving, and length of the dry period affect the likelihood of calves' infection with Cryptosporidium spp. considering these factors influence both colostrum and transition milk quality. The main results showed that 26.1% of calves were positive for the presence of Cryptosporidium spp. oocysts. The presence of clinical signs of diarrhea was recorded in 15% of the positive animals. Regression analysis showed that multiparous cows decrease the chance of calves to have Cryptosporidium spp. by 82%-89%, while cows calved on small farms decrease the chance of calves to have Cryptosporidium spp. by 80%. We suggest that primiparous cows are spending inner resources primarily on their maturation, thereby leaving the prerequisites for the infection of their offspring, while intense farming just increases the chance of unprotected calves to obtain infections.

Prevalence, risk factor and diversity of Cryptosporidium in cattle in Latvia.

The epidemiology of Cryptosporidium spp. in Latvia was investigated by testing fecal samples from 926 animals aged from one day to 24 years for the presence of Cryptosporidium spp. oocysts. The samples were collected from 87 cattle farms and from four slaughterhouses, and analyzed by conventional and fluorescent microscopy, followed by Cryptosporidium species and C. parvum subtype differentiation. Moreover, using a questionnaire, we surveyed factors that could be relevant as risk factors of Cryptosporidium spp. infection on the farms. Cryptosporidium spp. were shed by 33.8% of the investigated cattle and at least one shedding animal was found on 77.8% of the farms. In the present study, all four Cryptosporidium species reported to commonly infect cattle and two additional Cryptosporidium species (C. scrofarum and C. ubiquitum) were identified. In addition, mix infections of C. parvum/C. bovis, C. bovis/C. ryanae, C. parvum/C. ryanae, C. parvum/C. andersoni and C. bovis/C. andersoni were observed. C. parvum and C. bovis was mostly prevalent in young animals (0-3 months old) and in addition, diarrhea associated with C. parvum infection was observed only in very young animals. Cryptosporidium andersoni and C. ryanae in age group 0-3 months was observed in low prevalence, while a higher proportion of animals with diarrhea associated with C. andersoni infection was observed in very young animals and with C. ryanae in animals age group 4-24 months. Eight previously described C. parvum subtypes were observed. The majority of the subtypes were in the IIa subtype family, while one subtype was identified from the IId subtype family. The most common subtype was IIaA15G2R1, which was found in 34.2% of the C. parvum successfully subtyped samples. The probability of Cryptosporidium spp. associated diarrhea in cattle decreased significantly with the age of the animals and a prolonged period during which calves were fed with milk.

Traditional Latvian herbal medicinal plants used to treat parasite infections of small ruminants: A review.

Numerous treatment agents offering prophylaxis against livestock parasites are commercially available. However, because of increasing antiparasitic drug resistance, the increased popularity of environmentally friendly lifestyle choices, and organic farming, there is more demand for new alternatives to livestock anthelmintic control strategies and medications. It is important to develop antiparasitics that are safe, effective, inexpensive, and environmentally safe. Local, traditional herbal plants such as tansy, mugwort, wormwood, and heather may serve as treatments for intestinal parasites of sheep. This overview provides knowledge of traditional Latvian plants with antiparasitic activities to establish a database for further research to develop new herbal antiparasitic drugs.

Effect of immunoglobulin G concentration in dairy cow colostrum and calf blood serum on Cryptosporidium spp. invasion in calves.

AIM: The research aimed to test the association between the level of immunoglobulin G (IgG) in bovine colostrum and calf blood serum and to evaluate its relation to Cryptosporidium spp. invasion in calves. MATERIALS AND METHODS: Fresh colostrum and fecal specimens from cows (n=114) as well as blood and fecal specimens from newborn calves (n=114) were collected in the dairy cattle farm. Investigated calves were separated from their mothers directly after birth and received 2 L of colostrum in two separate feedings within the first 24 h. Blood samples were taken from calves at the age of 2 days. Coprological samples were taken from calves at the age of 1, 10, and 15 days. Both colostrum and fecal samples from cows were taken on the 1st day after calf birth. Rectal fecal samples were collected separately from each calf and cow into plastic bags. The collected calf serum samples and bovine colostrum samples were tested for bovine IgG by competitive enzyme-linked immunosorbent assay kit bovine Ig. To record oocysts of Cryptosporidium spp. in feces, the flotation method was used. Binomial logistic regression was performed to ascertain the effects of IgG in bovine colostrum and calf blood serum on the likelihood of Cryptosporidium spp. infection in calves. RESULTS: The concentration of IgG in bovine colostrum was higher (70.7±26.6 g/L, mean±standard deviation) than that in calf blood serum (13.2±6.1 g/L); the statistically significant difference was 57.4 g/L (95% confidence interval, 52.4-62.4), t (124.872)=22.536, p<0.001. Mann-Whitney's U-test showed a significant difference between samples collected on days 10 and 15 of the experiment (U=1944, z=2.330, p=0.020). The higher number of oocysts in calf feces was recorded on day 15 (median=6.5) compared to day 10 (median=4). The prevalence of calf infection from days 10 to 15 increased from 26.3 to 45.6% and was at least 3 times higher than in cows. A statistically significant positive correlation was recorded between IgG concentration of cow colostrum and calf blood serum (r (114)=0.414, p=0.001), whereas a correlation between the concentration of IgG and the intensity of Cryptosporidium spp. infection was not recorded (p>0.05). The logistic regression model was not statistically significant (χ2(2)=0.013, p=0.99 (10 days) and χ2(2)=0.100, p=0.95 (15 days)). CONCLUSION: Mother passive transfer of immunity to the offspring through colostrum does not influence the susceptibility of calves to Cryptosporidium infestation.

Review of Cryptosporidium and Giardia in the eastern part of Europe, 2016.

IntroductionThis paper reviews the current knowledge and understanding of Cryptosporidium spp. and Giardia spp. in humans, animals and the environment in 10 countries in the eastern part of Europe: Bosnia and Herzegovina, Croatia, Czech Republic, Estonia, Hungary, Latvia, Poland, Romania, Serbia and Slovenia. Methods: Published scientific papers and conference proceedings from the international and local literature, official national health service reports, national databases and doctoral theses in local languages were reviewed to provide an extensive overview on the epidemiology, diagnostics and research on these pathogens, as well as analyse knowledge gaps and areas for further research. Results:Cryptosporidium spp. and Giardia spp. were found to be common in eastern Europe, but the results from different countries are difficult to compare because of variations in reporting practices and detection methodologies used. Conclusion: Upgrading and making the diagnosis/detection procedures more uniform is recommended throughout the region. Public health authorities should actively work towards increasing reporting and standardising reporting practices as these prerequisites for the reported data to be valid and therefore necessary for appropriate control plans.

High prevalence of Trichinella spp. in sylvatic carnivore mammals of Latvia.

Trichinella spp. are zoonotic parasites transmitted to humans by the consumption of raw or insufficiently cooked meat of different animal species. Carnivore mammals are important reservoir hosts of these nematodes. The aims of this work were to establish the prevalence of Trichinella spp. and infection intensity in sylvatic carnivore mammals of Latvia, to identify the etiological agents at the species level and their circulation in the Latvian regions. From 2010 to 2014, muscle samples were collected from 1286 hunted animals (2 European badgers, 137 pine martens, 24 stone martens, 4 golden jackals, 394 raccoon dogs, 668 red foxes, 23 grey wolves, and 34 Eurasian lynxes). Trichinella spp. larvae were isolated by muscle digestion. Overall, 633 animals (49.2%; 95% CI 46.5%-52.0%) belonging to all the eight investigated species, tested positive for Trichinella spp. larvae. Trichinella britovi was the most common species (94.0%; 95% CI 91.7%-95.7%). Trichinella nativa was detected in 30 animals as single (6, 1.1%; 95% CI 0.4%-2.3%) or mixed infection (24, 4.4%; 95% CI 2.9%-6.4%) with T. britovi. Trichinella spiralis was detected in only three animals as mixed infection with T. britovi. The high prevalence of Trichinella spp. infection in sylvatic carnivore mammals suggests that they are good indicators for the risk assessment of Trichinella spp. in Latvia.

Efficacy against nematode and cestode infections and safety of a novel topical fipronil, (S)-methoprene, eprinomectin and praziquantel combination product in domestic cats under field conditions in Europe.

A novel topical combination product (BROADLINE(®), Merial) composed of fipronil, (S)-methoprene, eprinomectin and praziquantel was evaluated for safety and efficacy against nematode and cestode infections in domestic cats. The study comprised a multi-centre, positive control, blinded, field study, using a randomized block design based on order of presentation for allocation. In total 196 client-owned cats, confirmed as positive for naturally acquired infections of nematodes and/or cestodes by pre-treatment faecal examination, were studied in seven countries in Europe. Pre-treatment faecal examination revealed the presence of Toxocara, hookworm, Capillaria and/or spirurid nematode infections in 129, 73, 33 or 1 cat(s), respectively; infections with taeniid and Dipylidium cestodes were demonstrated in 39 and 17 cats, respectively. Cats were allocated randomly to one of two treatments in a ratio of 2, topical fipronil (8.3%, w/v), (S)-methoprene (10%, w/v), eprinomectin (0.4%, w/v) and praziquantel (8.3%, w/v) (BROADLINE(®), Merial; 130 cats); and 1, topical PROFENDER(®) Spot-On (Bayer; 66 cats) and treated once on Day 0. For evaluation of efficacy, two faecal samples were collected, one prior to treatment (Day -4 ± 4 days) and one at the end of the study (Day 14 ± 5 days). These were examined for fecal forms of nematode and cestode parasites. For evaluation of safety, cats were examined by a veterinarian before treatment and at the end of the study, and cat owners recorded the health status of their cats daily until the end of the study. For cats treated with Broadline(®), the efficacy was >99.9%, 100%, and 99.6% for Toxocara, hookworms, and Capillaria, respectively; and the efficacy was >99.9%, >99.9%, and 98.5%, respectively, for the cats treated with Profender(®) (p<0.001 for all nematodes and both treatments). Efficacy was 100% for both cestodes for both treatments (p<0.001). No treatment related adverse experiences were observed throughout the study. For both treatments, every cat that completed the study was given a safety score of 'excellent' for both local and systemic evaluations. The topical combination product of fipronil, (S)-methoprene, eprinomectin and praziquantel was shown to have an excellent safety profile and demonstrated high levels of efficacy when administered once as topical solution to cats infected with nematodes and cestodes under field conditions.