Insect live larvae as a new nutritional model in duck: effects on gut health.

BACKGROUND: This study aimed to evaluate the effects of Hermetia illucens (Black soldier fly-BSF) and Tenebrio molitor (Yellow mealworm-YMW) live larvae as a new nutritional model on duck's gut health, considering gut histomorphometry, mucin composition, cytokines transcription levels, and microbiota. A total of 126, 3-days-old, females Muscovy ducks were randomly allotted to three dietary treatments (6 replicates/treatment, 7 birds/pen): (i) C: basal diet; (ii) BSF: C + BSF live larvae; (iii) YMW: C + YMW live larvae. BSF and YMW live larvae were administered on top of the basal diet, based on the 5% of the expected daily feed intake. The live weight, average daily gain, average daily feed intake and feed conversion ratio were evaluated for the whole experimental period. On day 52, 12 ducks/treatment (2 birds/replicate) were slaughtered and samples of duodenum, jejunum, ileum, spleen, liver, thymus and bursa of Fabricius were collected for histomorphometry. Mucin composition was evaluated in the small intestine through histochemical staining while jejunal MUC-2 and cytokines transcription levels were evaluated by rt-qPCR. Cecal microbiota was also analyzed by means of 16 S rRNA gene sequencing. RESULTS: Birds' growth performance and histomorphometry were not influenced by diet, with a proximo-distal decreasing gradient from duodenum to ileum (p < 0.001), respecting the physiological gut development. Mucin staining intensity and MUC-2 gene expression did not vary among dietary treatments, even though mucin intensity increased from duodenum to ileum, according to normal gut mucus physiology (p < 0.001). Regarding local immune response, IL-6 was higher in YMW group when compared to the other groups (p = 0.009). Insect live larvae did not affect cecal microbiota diversity, but BSF and YMW groups showed a higher presence of Helicobacter, Elusimicrobium, and Succinatimonas and a lower abundance of Coriobacteriaceae and Phascolarctobacterium compared to C birds (p < 0.05). CONCLUSIONS: The use of BSF and YMW live larvae as new nutritional model did not impair gut development and mucin composition of Muscovy ducks, but slightly improved the intestinal immune status and the microbiota composition by enhancing regulatory cytokine IL-6 and by increasing minor Operational Taxonomic Units (OTUs) involved in short-chain fatty acids production.

[The implementation of family nursing in a municipality of the Provence of Bolzano]. / L'esperienza dell'introduzione dell'infermiere di famigliain un comune della Provincia di Bolzano.

. The implementation of Family Nursing in a municipality of the Provence of Bolzano. In the Province of Bolzano, 45 Family and Community Nurses have been trained to date with a 1250-hour course of which 400 hours of practical training. The working model adopted is the Calgary Assessment model, based on ithe relationship with families. The experience of the District of Bassa Atesina, which includes ten municipalities and about 40,000 inhabitants, is described, and in particular the assignment of three family nurses to a mountain municipality with a population of 1700 inhabitants, located with a considerable territorial dispersion. The FCNs cared for 48 families with tailored care plans. The model will be extended to other Districts.

Effect of Insect Live Larvae as Environmental Enrichment on Poultry Gut Health: Gut Mucin Composition, Microbiota and Local Immune Response Evaluation.

The aim of this study was to evaluate the effect of Hermetia illucens (HI) and Tenebrio molitor (TM) live larvae as environmental enrichment on the mucin composition, local immune response and microbiota of broilers. A total of 180 four-day-old male broiler chickens (Ross 308) were randomly allotted to three dietary treatments (six replicates/treatment; ten animals/replicate): (i) control (C); (ii) C+HI; (iii) C+TM. Live larvae were distributed based on 5% of the expected daily feed intake. At slaughter (39 days of age), samples of duodenum, jejunum and ileum (twelve animals/diet) were submitted to mucin histochemical evaluation. Expression of MUC-2 and cytokines was evaluated by rt-qPCR in jejunum. Mucin staining intensity was not influenced by diet (p > 0.05); however, this varied depending on the intestinal segment (p < 0.001). No significant differences were recorded for IL-4, IL-6 TNF-α, MUC-2 and INF-γ gene expression in jejunum, while IL-2 was lower in the TM group compared to HI and C (p = 0.044). Caecal microbiota showed higher abundance of Clostridium, Saccharibacteria and Victivallaceae in the HI group, while Collinsella was higher in the TM group. The results suggested that live insect larvae did not impair mucin composition or local immune response, and can slightly improve caecal microbiota by enhancing a minor fraction of short chain fatty acid-producing taxa.

Bronchopulmonary Nematodes in Alpine Ibex: Shedding of First Stage Larvae Analyzed at the Individual Host Level.

Pneumonia is the most frequent cause of death for Alpine ibex (Capra ibex) in Gran Paradiso National Park, (Italy). The etiology of this form of pneumonia is currently unknown and the identification of the primary etiological agent remains difficult due to biological and logistic constraints. Uncovering individual differences in Protostrongylid prevalence and intensity is important to further investigate the epidemiology of respiratory diseases and their relationship to heterozygosity and inbreeding in a once almost extinct population like C. ibex. In a group of 21 individually recognizable adult male we monitored monthly prevalence and intensity of Protostrongylid first-stage larvae using Baerman's technique from June to September 2019. First-stage larvae of 5 genera were detected. Muellerius (P = 100%, CI95% = 84-100) and Protostrongylus (P = 86%, CI95%:71-100) were two dominant genera according to Bush's importance index. Neostrongylus (P = 38%,CI95%: 17-59), Cystocaulus (P = 33%,CI95% = 13-53) were classified as co-dominant genera while Dictyocaulus filaria (P = 0.05%, CI95% = 0.04-0.13) was detected, for the first time in Alpine ibex, in one subject. Protostrongylidae larval excretion varied significantly over time, with minimum L1 excretion in July. Individual median larval intensity ranged from 4.4 lpg to 82.2 lpg with Poulin's discrepancy index showing highly aggregated distribution patterns for Muellerius spp. (D = 0.283, CI95% = 0.760-0.895) and Protostrongylus spp. (D = 0.635, CI95% = 0.580-0.705). Presented data provide the necessary base point to further investigate how lungworm infection account for the different rates of progression of pneumonia in C. ibex. Individual aggregation of larval intensity must be further evaluated to determine whether these differences mirror different levels of parasitic infection related to individual differences in immune response, hormonal-states or genetic fitness.

Reduced Efficacy of Fenbendazole and Pyrantel Pamoate Treatments against Intestinal Nematodes of Stud and Performance Horses.

Nematodes are an important cause of disease and loss of performance in horses. Changes in the parasitic fauna of horses have occurred in the past few decades, making cyathostomins the major parasites in adult horses, while large strongyles have become less prevalent. Parascaris spp. remains the most important parasite infecting foals and weanlings. Anthelmintic resistance is highly prevalent in cyathostomins and Parascaris spp. worldwide and it must be factored into treatment decisions. To assess anthelmintic efficacy in Northern Italy, we sampled 215 horses from 17 sport and horse-breeding farms. Fecal egg count reduction tests (FECRT) were used to assess anthelmintic efficacy. Copromicroscopic analysis was performed using MiniFLOTAC before treatment with fenbendazole, pyrantel pamoate or ivermectin, and repeated 14 days post-treatment. Strongyle-type eggs were detected in 66.91% of horses (CI95% 61.40-73.79%), while Parascaris spp. was detected in 2.79% (CI95% 1.94-5.95%). Reduced efficacy against cyathostomins was observed for fenbendazole in 55.56% of the treated animals (CI95% 41.18-69.06%), and for pyrantel pamoate in 75% of animals (CI95% 30.06-95.44%). Ground-based actions must be set in place to promote the uptake of state-of-the-art worm control plans that will prevent clinical disease while minimizing the selection pressure of resistant parasites.

A systematic review and meta-analysis of Toxoplasma gondii in roe deer (Capreolus capreolus) and red deer (Cervus elaphus) in Europe.

The objective of this systematic review and meta-analysis was to summarize the available knowledge on the seroprevalence of T. gondii in roe deer (Capreolus capreolus) and red deer (Cervus elaphus) in Europe. A computerized literature search of electronic databases (PubMed and CAB abstracts) was performed along with hand searches of library resources for relevant papers, books, abstracts and conference proceedings. A random-effect model was employed to calculate pooled seroprevalence estimates with 95% confidence intervals, and I2 statistic was used to assess heterogeneity. Further, moderator analysis was performed to evaluate the effect of geographical area on the seroprevalence in roe deer. From a total of 190 studies initially identified, 16 and 8 articles were included for roe deer and red deer, respectively. These comprise 3,913 roe deer and 2,913 red deer from different European countries. The pooled seroprevalence was estimated to be 29% (95% CI: 23%-35%) in roe deer and 15% (95% CI: 10%-20%) in red deer. High heterogeneity was detected in the seroprevalence data within each species. In roe deer, the pooled seroprevalence estimate was significantly different according to geographical area with 40% (95% CI: 31%-49%) in Western Europe, 31% (95% CI: 21%-43%) in Northern Europe, 27% (95% CI: 15%-41%) in Eastern Europe and 21% (95% CI: 14%-28%) in Southern Europe. The present study indicates a moderate exposure to T. gondii in roe deer and red deer in Europe, with very high prevalence in Western Europe. Our results highlight the significant risk associated to the consumption of venison, encouraging proper handling and cooking of game meat to prevent toxoplasmosis in humans.

Zoonotic Pathogens in Ticks from Migratory Birds, Italy.

Migratory birds can transport infected ticks across continents. We evaluated pathogens in ticks collected from migratory birds in Italy. We found DNA from Rickettsia aeschlimannii, R. africae, and R. raoultii bacteria, all of which can cause disease in humans. Bird migrations might facilitate the spread of these pathogens into new areas.

Ecological niche modeling of Babesia sp infection in wildlife experimentally evaluated in questing Ixodes ricinus.

Tick-borne diseases and especially protozoa of the genus Babesia, are gaining increasing attention as emerging zoonotic pathogens. Zoonotic species like B. venatorum and B. microti have wild animals as main reservoir hosts. We propose a habitat suitability model for Babesia spp., as tool for institutions and policy makes to better understand the entity of Babesia presence, to improve diagnostic awareness and to optimize screening and preventive actions. The probability of presence of Babesia spp. was estimated using as presence data, wild ruminants positive by PCR to Babesia spp. which were correlated to environmental factors that can favor or limit vector and host availability. We developed three separate models to discriminate the different roles of Red deer and Roe deer and Alpine chamois in Babesia spp. epidemiology. A comprehensive model using all presence data from all ungulates species  was also developed. The overall suitable area for Babesia spp. in this simulation is of 3723 km2, which correspond to 15.51% of the background regional territory. The model developed was empirically validated assessing tick abundance in randomly chosen areas classified by the model as moderately or highly suitable for Babesia spp. Collected ticks were tested by PCR for Babesia spp. to confirm model predictions as infection prevalence with Babesia spp. was significantly higher in areas predicted as highly suitable compared to those classified by the model as moderately suitable for Babesia spp.  (X2=5.05 p<0.05, Odds Ratio OR= 2.12 CI95% 1.1-4.1).

First detection and molecular identification of Anaplasma phagocytophilum in an introduced population of Reeve's muntjac (Muntiacus reevesi) in United Kingdom.

In the present study, we investigated blood samples of 196 invasive Reeve's muntjac (Muntiacus reevesi) and 91 native roe deer (Capreolus capreolus) originating from the same area in Thetford Forest in Eastern England for the occurrence of blood pathogens such as Anaplasmatacae, Rickettsiales and Piroplasmida (Babesia spp., Theileria spp.) by using PCR. Babesia spp., Rickettsia spp. and Theileria spp. were not detected. Only two male (1%) Reeve's muntjacs and six (6.6%) roe deer were positive for Anaplasma phagocytophilum with 100% identity among their sequences. However, it is not clear whether Reeve's muntjac is less susceptible to infection, less susceptible to infestation by I. ricinus, or an infection in Reeve's muntjac is more lethal and therefore less positive animals are taken during hunting events.

Molecular Survey on Vector-Borne Pathogens in Alpine Wild Carnivorans.

In Europe, free-ranging wildlife has been linked to the emergence of several vector-borne diseases such as rodents for Borrelia burgdorferi s.l. In particular, wild carnivorans are one of the most important sources of emerging zoonotic pathogens worldwide, although little information is available regarding the epidemiology of vector-borne parasites in these animals. Thus, the aim of this paper was to investigate the prevalence of Babesia spp., Anaplasma spp., Ehrlichia spp., Hepatozoon spp. and Leishmania infantum in alpine wild canids and mustelids from Italy. For this study, spleen samples of 157 foxes (Vulpes vulpes), 45 badgers (Meles meles), and 33 wolves (Canis lupus) collected between 2009 and 2017 in Northwest Italy were examined by using conventional PCR. Logistic regression was used to identify possible risk factors for pathogen infections. DNA of any of the tested pathogens was found in more than 90% of the analyzed animals. In particular, Babesia spp. showed significantly higher prevalence in foxes (89.7%) and badgers (89.6%) than in wolves, while the latter were considerably more infected with Hepatozoon canis (75.8%) than foxes (5.1%). None of the badger tested positive for Hepatozoon spp., although they showed high prevalence of Leishmania infantum (53.3%). Sequencing results revealed the presence, among others, of Babesia vulpes, Babesia sp. isolate badger type A and B, and Anaplasma phagocytophilum. Moreover, previously unreported pathogen/host associations were observed, such as Babesia capreoli in wolves and badgers. The prevalence of vector-borne pathogens observed in the present study is one of the highest reported so far, suggesting the importance of free-ranging carnivorans in the epidemiology and maintenance of the sylvatic cycle of the pathogens. Moreover, several of these pathogens are of particular importance regarding human (A. phagocytophilum, L. infantum) and pet health (L. infantum, B. vulpes).

Tick-borne pathogens in Ixodidae ticks collected from privately-owned dogs in Italy: a country-wide molecular survey.

BACKGROUND: Ticks and tick-borne diseases are increasingly recognized as a cause of disease in dogs worldwide. The epidemiology of ticks and tick-transmitted protozoa and bacteria has changed due to the spread of ticks to urban and peri-urban areas and the movement of infected animals, posing new risks for animals and humans. This countrywide study reports information on distribution and prevalence of pathogens in ticks collected from privately-owned dogs in Italy. We analyzed 2681 Ixodidae ticks, collected from 1454 pet dogs from Italy. Specific PCR protocols were used to detect i) Piroplasms of the genera Babesia and Theileria, ii) Gram-negative cocci of the family Anaplasmataceae and iii) Borrelia burgdorferi sensu lato. Sequencing of positive amplicons allowed for species identification. RESULTS: Babesia/Theileria spp. DNA was detected in 435 homogeneous tick-pools (Minimum Infection Rate (MIR) = 27.6%; 95% confidence interval (CI) = 25.4-29.8%) with higher prevalence in Ixodes ricinus and Rhipicephalus sanguneus group. The zoonotic B. venatorum was the most prevalent species (MIR = 7.5%; 95% CI = 6.3-9.0%). Anaplasma and Ehrlichia species were detected in 165 tick-pools (MIR = 10.5%; 95% CI = 9.3-11.8%) and specifically, A. phagocytophilum was identified with MIR = 5.1% (95% CI = 4.1-6.3%). Borrelia burgdorferi s.l. and B. afzelii were detected with MIR = 0.4% (95% CI = 0.2-0.8%) and MIR = 0.3% (95% CI 0.1-0.7%) respectively. CONCLUSIONS: Zoonotic pathogens B. venatorum and A. phagocytophilum were the most frequently detected in ticks collected from privately-owned dogs which might be used as markers of pathogens presence and distribution.

Absence of Viable Toxoplasma gondii in Artisanal Raw-Milk Ewe Cheese Derived from Naturally Infected Animals.

The presence of viable Toxoplasma gondii was investigated in artisanal cheeses made from milk of naturally infected ewes. Ewe milk was analyzed beforehand for the presence and vitality of T. gondii by loop-mediated isothermal amplification (LAMP) and reverse-transcriptase PCR (RT-PCR), respectively. Cheeses were prepared from raw milk following a traditional cheesemaking process. The cheese obtained from T. gondii-positive milk was analyzed by LAMP to detect Toxoplasma DNA-positive samples. RT-PCR was then carried out to assess the viability of the parasites in T. gondii-positive milk samples and fresh cheese, after 5 and 15 days of ripening. Physical-chemical parameters of cheeses were also investigated. All cheese samples derived from T. gondii-positive milk were positive according to LAMP, at both 5 and 15 days of ripening, while none of the samples were positive according to RT-PCR. Thus, while the presence of the parasite was demonstrated by the detection of specific DNA, the absence of detectable T. gondii RNA supports the hypothesis that changes in the chemical and physical characteristics occurring during the cheesemaking process and ripening period, could be sufficient to inactivate viable T. gondii in milk, minimizing the risk of human infection through consumption of raw sheep milk cheese.

Survey on tick-borne pathogens in ticks removed from humans in Northwestern Italy.

Ticks are able to transmit several pathogens to the host while feeding, and thus are considered the most important vectors of infectious agents together with mosquitos. The global incidence of tick-borne diseases (TBDs) is rising, due to increased interactions between pathogens, hosts and vectors, linked to global changes. Given that information about the prevalence of tick-borne pathogens in ticks removed from humans in Italy are scarce, the aim of the present study was to identify the species of ticks biting humans in Northwestern Italy and tick-borne pathogens they harbour. An overall number of 128 ticks from 92 patients were collected from April to October 2018, almost 98% of which belonging to the Ixodes ricinus species. Molecular analysis showed the presence of Babesia spp. in 29 out of 93 analysed tick pools, with a Minimum Infection Rate (MIR) of 31.18% (29/93; CI95% 22.67-41.19%), while 1 out of 93 pools tested positive for SFG Rickettsiae (MIR = 1.08%; CI95% 0.19-5.84%). No samples tested positive for A. phagocytophilum and Borrelia spp. Sequencing revealed the presence of Babesia venatorum (28 pools), Theileria buffeli/orientalis complex (1 pool) and Rickettsia monacensis. Among these, B. venatorum and R. monacensis are zoonotic pathogens able to cause from moderate to severe infections in humans. These data highlight the importance of passive surveillance to assess the epidemiology of TBDs that pose a threat to human health.

A national survey of Ixodidae ticks on privately owned dogs in Italy.

BACKGROUND: The geographical distribution of ticks on companion animals needs to be monitored to develop and plan effective control measures, as suggested by the European Scientific Counsel on Companion Animal Parasites. The aim of this study was to conduct the first Italian national survey of tick distribution on privately owned dogs. METHODS: The study was performed over 20 months (February 2016 - September 2017) and involved 153 veterinary practices in 64 different provinces covering 17/20 (85%) Italian regions. Participating practitioners were asked to examine five different dogs per month at random and complete a questionnaire for each dog. Differences in tick infestation associated with: sex, age and hair length (long and short); the dog's habitat (indoor or outdoor/kennel); and the dog's environment (urban or rural/sylvatic) were evaluated. The attachment site of ticks on the dog was also recorded. Acaricide efficacy was evaluated for the subset of dogs for which complete information on product used, date of sampling and date of last ectoparasiticide treatment was available. RESULTS: Of the 3026 dogs examined, 1383 (45.7%) were carrying at least one tick. Overall, 2439 tick samples were collected and a total of 14 tick species identified. Rhipicephalus sanguineus group were the most predominant ticks (63.6%), followed by Ixodes ricinus (30.6%) and I. hexagonus (5.6%). Twenty-four dogs had mixed tick infestations. Long-haired dogs had a higher tick infestation risk as did dogs with outdoor and rural/sylvatic lifestyles. Ticks were located on the head (37.4%), the neck (28.8%), the muzzle (15.5%) and the back (15.3%). A higher prevalence of Rhipicephalus was found in the interdigital spaces (10.8%) compared to Ixodes (0.2%). Finally, ectoparasiticide treatments were found significantly protective against tick infestation, especially orally administered formulations. CONCLUSIONS: Privately owned dogs in Italy have a high prevalence (45.7%) of infestation with ixodid ticks and this risk varies by dog phenotype and lifestyle.

Circulating genotypes of Toxoplasma gondii in Northwestern Italy.

Toxoplasma gondii is an apicomplexan parasite that in Europe is genetically characterized by three main clonal genotypes, with a lesser prevalence of atypical patterns. Data on the genotypes of T. gondii circulating both in wildlife and livestock in Northern Italy are scarce. In the present study skeletal muscle samples of cattle (Bos taurus), swine (Sus scrofa domesticus), fox (Vulpes vulpes), roe deer (Capreolus capreolus) and wild boar (Sus scrofa) were genotyped by using a nested PCR of 6 loci (alt.SAG2, GRA6, 5'SAG2, BTUB, C22-8 and SAG1) and in silico RFLP. High prevalence of genotype I and non-canonical genotypes were observed in this study, with some differences in the population structure of the parasite between livestock and wildlife. Genetic variability of T. gondii in Europe could be more variable than previously thought, with possible implication for public health.