First Report of Isolation and Characterization of Acanthamoeba spp. from the Milk Used for Calf Feeding.

PURPOSE: Acanthamoeba spp. can be found in natural and artificial environments, which reflects their high adaptability to different conditions. Based on the available data, there is scarce information about the isolation of amoeba from milk. This study aimed to investigate the probable presence of Acanthamoeba in milk used for calf feeding. METHODS: 200 milk samples from 50 industrial and traditional farms were collected. The samples were filtered and cultured on the 1.5% Non-nutrient agar medium. The amoebic growth was examined with an inverted microscope daily. DNA was extracted from the positive plates, and a PCR reaction was undertaken using the primers amplifying the Acanthamoeba 18 S rRNA gene. Five samples were purified and sequenced using specific primers. Maximum likelihood reconstructions were performed using the phylogenetic program MEGA software. The osmo and thermotolerance of isolated trophozoites were examined as well. RESULTS: Out of 200 milk samples, Acanthamoeba was isolated from 27 (13.5%). The phylogenetic tree represents that all the isolates belonged to the genotype T4. Results of thermo and osmotolerance tests showed that isolates could develop at 37 and 43 ◦C. Besides, trophozoites survived at 0.5 M mannitol and 1 M. CONCLUSION: For the first time, Acanthamoeba spp. were isolated from milk used to feed dairy calves. Due to Acanthamoeba's neglected role in pathogen persistence and survival, hygiene instructions should be reconsidered.

Compatibility, Cytotoxicity, and Gastrointestinal Tenacity of Bacteriocin-Producing Bacteria Selected for a Consortium Probiotic Formulation to Be Used in Livestock Feed.

Bacteriocin-producing Escherichia coli ICVB442, E. coli ICVB443, Enterococcus faecalis ICVB497, E. faecalis ICVB501, and Pediococcus pentosaceus ICVB491 strains were examined for their pathogenic risks and compatibility and hence suitability as consortium probiotic bacteria. Except for E. coli ICVB442, all were inclined to form biofilm. All were gelatinase-negative, sensitive to most of the antibiotics tested and not cytotoxic to porcine intestinal epithelial cells (IPEC-1) when tested at a multiplicity of infection (MOI) of 1. P. pentosaceus ICVB491 stood apart by inhibiting the other four strains. Both E. coli strains and E. faecalis ICVB497 strain were ß-hemolytic. Survival in the TIM-1 dynamic model of the human digestive system was 139% for the tested E. coli ICVB443 strain, 46% for P. pentosaceus ICVB491, and 32% for the preferred E. faecalis ICVB501 strain. These three potential probiotics, which are bacteriocin-producing strains, will be considered for simultaneous use as consortium with synergistic interactions in vivo on animal model.

Fatal Aberrant Parasite Migration of Echinuria uncinata in Two African Pygmy Geese (Nettapus auritus).

Two adult African pygmy geese (Nettapus auritus) were found dead with minimal clinical disease signs. Necropsy revealed aberrant migration of the nematode parasite Echinuria uncinata, as confirmed morphologically and through DNA sequencing. This common waterfowl parasite typically lives in the proventriculus, burying headfirst into the mucosa and laying eggs into the gastrointestinal lumen. In these geese, the parasites tunneled through the gastrointestinal tract wall to invade the coelomic cavity; from which, a substantial quantity of eggs found their way into the coelomic space and into the air sacs and lungs. This potential parasite migration should be monitored for in Anseriformes species that present with similar disease conditions, and the use of Daphnia species, the intermediate host, as a waterfowl feed source is not recommended.

Gastric canthariasis caused by invasion of mealworm beetle larvae in weaned pigs in large-scale farming.

BACKGROUND: Mealworm beetle T. molitor (Coleoptera: Tenebrionidae) (Linnaeus, 1758) is one of the most important cosmopolitan primary storage pests, scavenging on a variety of post-harvest grains and affecting the quality and safety of food and feed. In addition to being an important factor in feed hygiene, the insect can also be an epidemiological factor of canthariasis. Livestock infestations with T. molitor are rarely reported. This article describes T. molitor-caused canthariasis in pigs in large scale closed-cycle farming. RESULTS: In the spring, we registered a significantly increased mortality among weaned pigs. In autopsy, live 3-6 mm long T. molitor larvae were found in their stomachs, especially in the non-glandular oesophageal region, on average 2-3 larvae per 10 cm2 of gastric mucosa. Corrective actions reduced the number of deaths back to basal levels. CONCLUSIONS: This is the first documented case of potentially lethal gastric canthariasis in weaned pigs, caused by invasion of T. molitor larvae. Although canthariasis caused by T. molitor has not been a significant problem in farm animals so far, our case indicates that the presence of mealworm beetles is a potential threat to animal welfare and health.

Epidemiological survey of Toxoplasma gondii and Neospora caninum infections in dairy goats in Central-Southern Taiwan.

Toxoplasma gondii and Neospora caninum are intracellular protozoan parasites that cause reproductive disorders in ruminants and humans. Information on the risk factors of T. gondii and N. caninum infections in goats is very limited in Taiwan. The aim of the study was to investigate the epidemiology and identify the risk factors of these two infections in goats. A total of 630 caprine sera were collected from 42 dairy goat farms and the owners were interviewed by a structured questionnaire. The apparent seroprevalences of T. gondii in farm- and individual- levels were respectively 88.1% and 32.22%, while those of N. caninum were 19.05% and 2.54%, respectively. Toxoplasma gondii B1 gene was identified in 7 feed samples and 8 from the water samples whereas N. caninum was not found. Wooden flooring was the main risk factor for T. gondii infection while the frequency of visits by staff to other farms and the breed of goat were risk factors for N. caninum. The improvement of flooring materials or thorough cleaning, periodic disinfection and maintenance of dryness on the floor are highly recommended for the prevention of T. gondii infection in farmed goats. In addition, unnecessary visits to other farms should be limited to prevent the spread of N. caninum. These factors should be highlighted for the prevention of T. gondii and N. caninum in goats, particularly when raised in intensive housing system with flooring on height.

Critically Appraised Topic on Adverse Food Reactions of Companion Animals (8): Storage Mites in Commercial Pet foods.

BACKGROUND: Dogs with year-round atopic dermatitis are often sensitized to Dermatophagoides house dust mites (HDM). Storage mites (SM) are known to grow on cereal-rich foods. Tyrophagus SM can exacerbate clinical signs of allergy in laboratory dogs sensitized to HDM. Consequently, atopic dogs with high-levels of HDM-specific IgE are likely to have a flare of signs after eating a food contaminated with SM; the development of such flares would lead to a false positive diagnosis of food allergy. Herein, we reviewed the published evidence about the growth of SM on commercial dry pet foods. RESULTS: We searched two databases on January 25, 2019 for articles providing original information on the growth of SM on commercial dog foods. We found ten articles, five reporting results of laboratory experiments and five from field studies. Storage mites, especially Tyrophagus putrescentiae, can multiply on protein- and fat-rich dog foods. The population growth is higher when the initial mite density is high and when kibbles are crushed. When storage conditions lead to the overgrowth of molds on the kibbles, the mite proliferation is higher. Storage mites do not bore holes in food packages but invade bags via defective seals. In the field, SM contamination usually is undetectable in newly-opened commercial dog foods, and, if present, their number is low. When newly-purchased bags are stored in temperate conditions indoors, little overgrowth-if any-of SM occurs. However, when kept in environmental conditions with higher temperature and humidity, Tyrophagus mites will enter and proliferate in sealed food packages. CONCLUSIONS: Commercial dry pet foods should be kept indoors and sealed to decrease the risk of contamination with SM. When performing dietary restriction (elimination) and provocation trials for the diagnosis of food allergies in dogs, it seems preferable to choose newly-purchased bags-of both original and testing diets-to reduce the probability of their contamination with SM, especially Tyrophagus putrescentiae. In case of doubt about the presence of SM in any of these foods, one should perform food challenges with single home-cooked ingredients. Storage mite contamination might lead to an erroneous diagnosis of food allergy in HDM-sensitized dogs.

Risk Evaluation of Passive Transmission of Animal Parasites by Feeding of Black Soldier Fly (Hermetia illucens) Larvae and Prepupae.

HIGHLIGHTS: Black soldier fly larval intestine extracts do not affect coccidian oocysts or nematode eggs. Studied parasites passed through black soldier fly larvae or attached to the larval surface. Black soldier fly larvae as animal feed may pose a risk of parasite transmission. Larval washing is not sufficient for removing parasites; further hygiene steps are needed.

Goats worm burden variability also results from non-homogeneous larval intake.

For small ruminants, Gastrointestinal Nematodes (GINs) are responsible for severe economic losses and they are also an animal welfare problem. GIN use their host to reproduce and disperse eggs on the pasture, from where they can re-infect another animal. The high density of hosts on the pasture and the extreme tolerance of GIN to environmental constraints make GIN eradication almost impossible. In addition, significant resistance to anthelmintic treatment requires sustainable and integrated management to maintain the health and financial well-being of livestock farming. In this context, models of the complex interactions between host, GIN and environment can help us to design long term optimal management strategies. To build such models, quantitative information is needed but are generally very challenging to collect. In this article, we focus on the number of ingested larvae per animal, which we propose to characterise by using a simulation framework based on the estimation of the spatial distribution of the host over time. Our framework allows us to show that worm burden individual variation is not only explained by the host's genetics, as is often the case, but is also a result of the grazing spatial process.

Biochemical Properties of α-Amylase from Midgut of Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) Larvae.

The lesser mealworm, Alphitobius diaperinus (Panzer), is the main insect pest in the poultry industry, thus causing serious damage to production. In this work, the properties of midgut α-amylase from larvae of A. diaperinus were characterized, and its in vitro activity to proteinaceous preparations from different cultivars of common bean (Phaseolus vulgaris) was determined, as well as the amylolitic activity of insects reared on different types of poultry diet. In order to establish some assay conditions, time course and enzyme concentration upon the reaction rate were determined. Product proceeded linearly with time, and the activity was directly proportional to the enzyme concentration. Banding patterns in mildly denaturing electrophoresis showed a single band with apparent molecular weight of 42 kDa. α-Amylase reached optimal temperature at 45°C and pH 5.0 as the optimal one. It maintained 34.6% of the activity after being kept at 60°C for 5 min, and 23%, after 60 min. However, at 80°C, only 14 and 6% remained after 5 and 60 min, respectively. The presence of Ca2+ and Na+ ions decreased the enzyme activity at concentrations higher than 2 and 100 mM, respectively. The activity was significantly inhibited by some proteinaceous extracts from common bean cultivars, and it declined with increasing proteinaceous concentration. No significant difference was observed when the amylolytic activity was determined in A. diaperinus reared on different poultry diets, offered to broilers in the starter, grower, finisher, and layer phases.

Persistence of Trichomonas gallinae in Birdseed.

Trichomonas gallinae has emerged worldwide as a cause of mortality in songbirds (passerines). The congregation of numerous birds, including the reservoir hosts, pigeons and doves (columbids), at backyard feeding and watering sources has been suggested as a potential driver for the outbreaks. Evidence supporting a role for water in transmission has been established, but the role of birdseed in the transmission of trichomoniasis remained to be investigated. We assessed the survival of T. gallinae in three commercial birdseeds (mixed seed, black-oil sunflower seed, and niger seed) routinely used to attract passerine birds to local properties. Trichomonad suspensions were inoculated (low dose: 1 × 103; high dose: 1 × 105) into each of the three seed types in petri dishes, using both dry and moist (water-soaked) conditions, in triplicate. Petri dishes were incubated at 37 C and monitored for T. gallinae survival for 48 hr by wet-mount microscopy and by InPouch™ TF medium culture for 10 days. Surviving trichomonads were not detected in any of the dry birdseed treatments. In moist conditions, however, trichomonads were found to survive ≤24 hr in all three seed types and ≤48 hr in the mixed seed that contained organic debris. We demonstrate that T. gallinae has the ability to survive in moist birdseed, which suggests that public bird-feeding sites may play a significant role in the transmission of trichomoniasis.

The impact of socio-cultural factors on transmission of Taenia spp. and Echinococcus granulosus in Kosovo.

Echinococcus granulosus sensu latu (s.l.) and Taenia hydatigena are common parasites of ruminant intermediate hosts in the Balkans. Transmission is linked mainly to home slaughtering and the feeding of infected organs to dogs. In Kosovo, many old sheep are slaughtered particularly during Eid al-Adha (Feast of Sacrifice). To determine whether this tradition could affect parasite transmission, we compared the probability of 504 dogs to contract taenid infections after deworming during one period before Eid al-Adha and a similar period beginning with this event. Initially, taeniid eggs were detected in 6·2% (CI 4·2-8·6) of the dogs. The prevalence before Eid al-Adha was significantly lower (1·2%, CI 0·4-2·6) as compared with the prevalence after the event (4·3%, CI 2·6-6·3). A comparable trend was apparent at species level for T. hydatigena and E. granulosus. These results indicate that the pronounced increase of taeniid infections, including E. granulosus s.l., after Eid al-Adha is linked to traditional home slaughtering that occurs during this celebration. This particular epidemiological situation provides an opportunity for implementing focussed control activities.

Infection with Paragonimus westermani of boar-hunting dogs in Western Japan maintained via artificial feeding with wild boar meat by hunters.

Infection of boar-hunting dogs with Paragonimus westermani was investigated in Western Japan. Blood and rectal feces were collected from 441 dogs in the three districts (205 in Kinki, 131 in Chugoku and 105 in Shikoku District). In a screening ELISA for serum antibody against P. westermani antigen, 195 dogs (44.2%) showed positive reaction. In the 195 dogs, 8 dogs were found excreting P. westermani eggs after molecular analysis of fecal eggs, and additional 7 were identified serologically for the parasite infection because of their stronger reactivity against P. westermani antigen than against antigens of other species of Paragonimus. A spatial analysis showed that all of the P. westermani infections were found in Kinki and Chugoku Districts. In this area, dogs' experience of being fed with raw boar meat showed high odds ratio (3.35) to the sero-positivity in the screening ELISA, and the frequency of such experiences was significantly higher in sero-positive dogs. While clear relationship was not obtained between predation of boars by dogs during hunting and their sero-positivity. Therefore, it is suggested that human activity of feeding with wild boar meat is the risk factor for P. westermani infection in boar-hunting dogs. Considering that hunting dogs could play as a major definitive host and maintain the present distribution of P. westermani in Western Japan, control measures for the infection in hunting dogs, such as prohibition of raw meat feeding and regular deworming, should be undertaken.

Coprodiagnosis of Echinococcus granulosus infection in dogs from Ankara, Turkey.

The present study were undertaken to compare two isolation techniques (centrifugal flotation and sedimentation) for recovering taeniid eggs from faecal samples, to identify E. granulosus DNA from taeniid eggs by PCR, and to determine the prevalence of E. granulosus in dogs in villages. Faecal samples were collected from 100 dogs in Ankara province. Taenia spp. eggs were found in 27% of dogs faeces. Echinococcus granulosus-specific PCR was obtained in 14 (51.85%) of the taeniid eggs-positive samples. As well as finding Taenia eggs in dogs' faeces, we also found eggs of some helminthic parasites; such as Dipylidium caninum, Toxocara canis, Toxascaris leonina, Trichuris sp., Capillaria sp., Filaroides sp., Dioctophyme renale, Linguatula serrata, hookworm, Dicrocoelium sp., Fasciola sp. and Ascaridia galli. Significantly, more dogs excreting taeniid eggs were diagnosed with the sedimentation method (n=27) as compared to the flotation method (n=10).

Nutritional composition of black soldier fly (Hermetia illucens) prepupae reared on different organic waste substrates.

BACKGROUND: Black soldier fly larvae are converters of organic waste into edible biomass, of which the composition may depend on the substrate. In this study, larvae were grown on four substrates: chicken feed, vegetable waste, biogas digestate, and restaurant waste. Samples of prepupae and substrates were freeze-dried and proximate, amino acid, fatty acid and mineral analyses were performed. RESULTS: Protein content of prepupae varied between 399 and 431 g kg-1 dry matter (DM) among treatments. Differences in amino acid profile of prepupae were small. On the other hand, the ether extract (EE) and ash contents differed substantially. Prepupae reared on digestate were low in EE and high in ash (218 and 197 g kg-1 DM, respectively) compared to those reared on vegetable waste (371 and 96 g kg-1 DM, respectively), chicken feed (336 and 100 g kg-1 DM, respectively) and restaurant waste (386 and 27 g kg-1 DM, respectively). Prepupal fatty acid profiles were characterised by high levels of C12:0 in all treatments. CONCLUSION: Since protein content and quality were high and comparable for prepupae reared on different substrates, black soldier fly could be an interesting protein source for animal feeds. However, differences in EE and ash content as a function of substrate should be considered. © 2016 Society of Chemical Industry.

A new species of Thyreophagus (Acari: Acaridae) from Brazil, with notes on species associated with stored food and human habitats and a key to species of this genus.

A new species Thyreophagus cracentiseta n. sp. is described morphologically based on adult females, adult homeomorphic and heteromorphic males collected from chicken feed in Brazil. The Thyreophagus species associated with stored food and human habitats are reviewed, and a key to separate species of this genus is provided.

[Tyrophagus palmarum and its hypopus found in feed of Tenebrio molitor].

OBJECTIVE: To observe the Tyrophagus palmarum and its hypopus in feed of Tenebrio molitor. METHODS: Feed samples were collected from T. molitor farms, and T. palmarum and its hypopus were separated from the samples by the vibration sieve method and floating method. The glass specimens were prepared and T. palmarum and its hypopus were observed under an optical microscope. RESULTS: Under the light microscope, the skin of T. palmarum was smooth and bright, the color of legs and chelicerae was relatively dark, the length of dorsal seta d2 was 3-4 times longer than d1 and la; The ω1 in tarsus of leg Ⅰ, Ⅱ was short rod, and its top was not tapered; the sucker plate in tarsus of leg Ⅳ located in the middle of the section. The hypopus was oblate, and there were two pairs of legs in front which extended to body. The rear end of hypopus became pointless and round, wrapping around a transparent and ossification skin. CONCLUSIONS: Light microscopy shows the morphological characteristics of T. palmarum and its dormant body, providing the basis for identifying the hypopus.

Diphyllobothrium sp. in Canis familiaris from the subtropical area of Argentina (Puerto Iguazú, Misiones).

This paper reports the first finding of Diphyllobothrium sp. eggs in Canis familiaris (domestic dog) from Puerto Iguazú, a subtropical city of Misiones province, Argentina. In 2013, two positive cases of Diphyllobothrium sp. eggs were detected during an annual parasitological survey of dogs. Dog feces were collected in vials containing 10% formalin and processed using Telemann's sedimentation and Sheather's flotation techniques. The two cases were detected in rural areas of the municipality. Since Misiones is not a part of the endemic area of diphyllobothriasis and given the fact that it is located in the three-border area of Argentina, Brazil and Paraguay, we consider this finding of great importance to public health. We stress the need for updating the current knowledge about the life cycle of these parasites considering the range of intermediate and definitive hosts, their zoonotic potential, and the epidemiological situation in non-endemic areas.

Duddingtonia flagrans chlamydospores in nutritional pellets: effect of storage time and conditions on the trapping ability against Haemonchus contortus larvae.

The study evaluated the effect of storage time and conditions of nutritional pellets (NP) containing Duddingtonia flagrans chlamydospores on its in vitro trapping ability against Haemonchus contortus L3 larvae. The treated batch (200 NP) contained 4 × 106 chlamydospores of the FTH0-8 strain, whereas the control batch (200 NP) was produced without spores. Both NP batches were exposed to four experimental storage conditions: (T1) shelves (indoors); (T2) refrigeration (4°C); (T3) outdoors under a roof; and (T4) 100% outdoors. Each group comprised 48 NP with spores and 48 NP without spores (control). The ability of D. flagrans spores to trap H. contortus L3 larvae was evaluated for 8 weeks for each storage condition. For that purpose, six randomly selected NP with spores were compared to their respective control NP. Each NP was individually crushed. The crushed material (1 g) was placed on the surface of a 2% water agar plate with 200 H. contortus L3 larvae. Plates were sealed and were incubated at room temperature for 8 days. The whole content of every plate was transferred to a Baermann apparatus to recover the remaining larvae. There was a clear larval reduction in the NP with spores, compared to the respective control NP in the four storage conditions (P< 0.05). The mean reductions ( ± SEM) of the storage conditions were 67 ± 4.9 (T2), 77 ± 6.1 (T1), 81.5 ± 3.8 (T4) and 82.1 ± 2.5 (T3). Larval reductions were similar at all times and were not affected by storage conditions or storage time (R 20.05). The long-term shelf-life of the chlamydospores in the NP suggests that this spore dosage technology is a viable option.

Pilot project to investigate over-wintering of free-living gastrointestinal nematode larvae of sheep in Ontario, Canada.

This study investigated the overwintering survival and infectivity of free-living gastrointestinal nematode (GIN) stages on pasture. The presence of GIN larvae was assessed on 3 sheep farms in Ontario with a reported history of clinical haemonchosis, by collecting monthly pasture samples over the winter months of 2009/2010. The infectivity of GIN larvae on spring pastures was evaluated using 16 tracer lambs. Air and soil temperature and moisture were recorded hourly. Free-living stages of Trichostrongylus spp. and Nematodirus spp. were isolated from herbage samples. Gastrointestinal nematodes were recovered from all tracer lambs on all farms; Teladorsagia sp. was the predominant species. Very low levels of Haemonchus contortus were recovered from 1 animal on 1 farm. The results suggest that Haemonchus larvae do not survive well on pasture, while Teladorsagia sp., Trichostrongylus spp. and Nematodirus spp. are able to overwinter on pasture in Ontario and are still infective for sheep in the spring.

Projet pilote pour faire enquête sur l'hivernage des larves de nématodes gastro-intestinaux libres chez les moutons en Ontario, au Canada. Cette étude a examiné la survie à l'hivernage et le pouvoir infectieux des stades des nématodes gastro-intestinaux (NGI) libres dans les pâturages. La présence de larves de NGI a été évaluée en recueillant des échantillons mensuels dans le pâturage pendant les mois de l'hiver 2009­2010 dans 3 fermes ovines en Ontario avec des antécédents documentés d'hémonchose clinique, tandis que le pouvoir infectieux des larves de NGI sur les pâturages du printemps a été évaluée en utilisant 16 agneaux sentinelles. La température et l'humidité de l'air et du sol ont été notées toutes les heures. Les stades libres de Trichostrongylus spp. et de Nematodirus spp. ont été isolés d'échantillons d'herbage. Les NGI ont été récupérés de tous les agneaux sentinelles dans toutes les fermes et Teladorsagia sp. était l'espèce prédominante. De très faibles taux d'Haemonchus contortus ont été récupérés chez 1 animal dans 1 ferme. Les résultats suggèrent que les larves d'Haemonchus ne survivent pas bien dans le pâturage, tandis que Teladorsagia sp., Trichostrongylus spp. et Nematodirus spp. peuvent survivre l'hiver dans le pâturage de l'Ontario et être toujours infectieux pour les moutons au printemps.(Traduit par Isabelle Vallières).

Investigating the relationship between feed and helminthic burden of captive birds of prey in Hong Kong.

The life cycle of most parasitic helminthes is related to their hosts feeding habits. Thus we need to investigate the impact of diet on the host's helminthic parasite burden. Not many studies in captive raptors have been conducted and published regarding parasitic infections. The aim of this study is to evaluate the helminthic burden of raptors kept in captivity and establish a relationship with the feed provided. A total of N = 51 different species of captive birds of prey were fed different diets consisting in different combinations of day old chicks, chicken breast, whole chicken carcass and mice. Their feces were sampled and the parasite burden was determined. A negative binomial model was successfully fitted to the data and the feeds "mice" (P < 0.001) and "whole chicken carcass" (P < 0.001) significantly contributed to an increase in the observed burden. Significant differences were also found between species (P < 0.001). Raptors fed adult animal carcasses and offal may explain the increase in the observed burden as these feeds have a larger probability of being contaminated by a larger variety of helminthic fauna.