Investigating on Besnoitia besnoiti (Apicomplexa, Sarcocystidae) in naturally infected dairy cattle by an integrated approach.

Bovine besnoitiosis, caused by Besnoitia besnoiti, is a (re)emerging disease in Europe (Cortes et al., 2014), including Italy (Gazzonis et al., 2014; 2017). However, its economic impact is scarcely considered and generally underestimated and there are still little studied aspects concerning both the parasite and the disease. Following a natural outbreak of besnoitiosis in a dairy herd, a study was planned to characterize B. besnoiti infection in cattle through a multidisciplinary approach.Suspicious abortions and clinical cases of besnoitiosis were reported in a dairy farm (September 2017, Northern Italy) housing 216 Holstein cattle. Blood samples were collected; haematological and serological analyses (ELISA and confirmatory WB) were performed (Fernandez-Garcia et al., 2009). Histology and molecular (endpoint ITS-1 PCR (Cortes et al., 2007) and sequencing) analyses of tissues from a slaughtered cow with chronic besnoitiosis were carried out. Out of 59 ELISA-positive animals, 50 (23%) were confirmed by WB. B. besnoiti prevalence was higher in cows (41%) than in calves (12%); any heifer did not result positive. Considering haematological parameters, a significant shift in the differential leucocyte formula from lymphocyte to granulocyte was recorded in infected cows (Mean±S.D.:L=46.1±18.4,G=53.9±18.4) if compared to negative animals (Student’s T-test,p=0.012). This finding could be helpful in diagnosis, treatment and control of besnoitiosis. Histology revealed a high load of B. besnoiti tissue cysts in skin, vulva, muzzle, sclera, eyelid, respiratory tract, emphasizing the possibility of parasite transmission through direct contact among animals. B. besnoiti was confirmed by PCR in other organs (heart, liver, aorta wall, tonsil) and especially in ovary, uterus and vulva, suggesting that the infection could affect cows’ fertility. Parasite DNA was also found in masseters posing an important question for food security, even if B. besnoiti is not considered zoonotic. The study suggests that to investigate the dynamics of bovine besnoitiosis is mandatory associate clinical and laboratory tests, including the genetic characterization of the parasite and its eventual correlation with the disease outcome

In vitro effects of different levels of quebracho and chestnut tannins on rumen methane production, fermentation parameters, and microbiota

Both condensed and hydrolysable tannins (CTs and HTs, respectively) have the ability to reduce enteric CH4 production in ruminants. However, the precise mechanism of action is not fully understood. Among the proposed hypotheses are the reduction of ruminal digestibility, direct control action on protozoa, reduction of archaea, and a hydrogen sink mechanism. In this in vitro study, which simulated rumen fermentation, two additives, one containing CTs (70% based on DM) from quebracho and one with HTs (75% based on DM) from chestnut, at four levels of inclusion (2, 4, 6, 8% on an as-fed basis) were added to the fermentation substrate and tested against a negative control. Both types of tannins significantly reduced total gas (GP) and CH4 (ml/g DM) production during the 48 h of incubation. The lower GP and CH4 production levels were linked to the reduction in dry matter digestibility caused by CTs and HTs. Conversely, no significant differences were observed for the protozoan and archaeal populations, suggesting a low direct effect of tannins on these rumen microorganisms in vitro. However, both types of tannins had negative correlations for the families Bacteroidales_BS11 and F082 and positive correlations for the genera Prevotella and Succinivibrio. Regarding the fermentation parameters, no differences were observed for pH and total volatile fatty acid production, while both CTs and HTs linearly reduced the NH3 content. CTs from quebracho were more effective in reducing CH4 production than HTs from chestnut. However, for both types of tannins, the reduction in CH4 production was always associated with a lower digestibility without any changes in archaea or protozoa. Due to the high variability of tannins, further studies investigating the chemical structure of the compounds and their mechanisms of action are needed to understand the different results reported in the literature

Toxoplasma gondii in small ruminants in Northern Italy – prevalence and risk factors

[b]Objective[/b]. The aim of the survey was to evaluate [i]Toxoplasma gondii[/i] seroprevalence in small ruminants and possible risk factors associated with the infection. [b]Materials and methods[/b]. Sera from 474 goats and 502 sheep reared on 42 farms in northern Italy were collected and tested for IgG antibodies to [i]T. gondii[/i] by IFAT (indirect immunofluorescence antibody test). To identify risk factors, a binary logistic regression analysis of the variables was performed. An audit form about farm management was used. [b]Results[/b]. Antibodies to [i]T. gondii [/i]were found in 96.6% of goat farms and in 87.5% of sheep farms; 41.7% goats and 59.3% sheep resulted positive. Seroprevalence was significantly higher in sheep than in goats. Seroprevalence values were similar in goats from eastern and western areas, whereas goats from the southern area were at lower risk of infection. Saanen goats presented the lowest seroprevalence (30.7 %), whereas cross-breed exhibited the highest rate (48.7%). Goats from farms housing both sheep and goats had an infection risk 1.39 times higher than goats from farms that did not house sheep. Animals bred on intensive farms showed lower prevalence (22.1%) in comparison with those from extensive (45.6%) or semi-intensive farms (60%). Sampling area was one of the strongest predictors of [i]T. gondii [/i]infection in sheep flocks. Transhumant flocks showed a higher risk of infection by [i]T. gondii[/i] compared with semi-intensive farms (66.8% vs. 38.4%). [b]Conclusions[/b]. The highest [i]T. gondii[/i] seroprevalence values were registered in transhumant flocks of sheep and in family businesses rearing goats. As these traditional activities represent an important resource for the conservation of the territory and its economy, management practices for a better control of the disease should be improved

Gastrointestinal nematodes of goats: host–parasite relationship differences in breeds at summer mountain pasture in northern Italy

The Orobic goat is a hardy breed native to the Orobic Alps (Lombardy, northern Italy). The aim of the study was the assessment of gastrointestinal nematode (GIN) egg excretion in Alpine and Saanen (cosmopolite breeds) and Orobic grazing goats, after a strategic treatment with eprinomectin in late June

Canine fecal contamination in a metropolitan area (Milan, north-western Italy): prevalence of intestinal parasites and evaluation of health risks

Intestinal parasites of dogs represent a serious threat to human health due to their zoonotic potential. Thus, metropolitan areas presenting high concentrations of pets and urban fecal contamination on public areas are at sanitary risk. Major aim of this survey was to determine prevalence of zoonotic parasites in dog fecal samples collected from public soil of Milan (north-western Italy). Differences in parasites prevalence distribution were explored by a geographical information system- (GIS-) based approach, and risk factors (human density, sizes of green parks, and dog areas) were considered. The metropolitan area was divided into 157 rectangular subareas and sampling was performed following a 1-kilometer straight transect. A total of 463 fecal samples were analyzed using centrifugation-flotation technique and ELISA to detect Giardia and Cryptosporidium coproantigens. A widespread fecal contamination of soil was highlighted, being fecal samples found in 86.8% of the subareas considered. The overall prevalence of intestinal parasites was 16.63%. Zoonotic parasites were found, such as Trichuris vulpis (3.67%), Toxocara canis (1.72%), Strongyloides stercoralis (0.86%), Ancylostomatidae (0.43%), and Dipylidium caninum (0.43%). Giardia duodenalis was the most prevalent zoonotic protozoa (11.06%), followed by Cryptosporidium (1.10%). Faeces from subareas characterized by broad green areas showed to be particularly prone to infection

Toxoplasma gondii infection and biosecurity levels in fattening pigs and sows: serological and molecular epidemiology in the intensive pig industry (Lombardy, Northern Italy)

Toxoplasma gondii is a widespread protozoan affecting animals and humans. One of the major routes of human infection is through the consumption of raw or under-cooked meat, particularly of certain animal species, including pigs. Although T. gondii represents an important public health issue, its control at slaughter is not mandatory. Consequently, available information on T. gondii infection in domestic animals destined for human consumption is scarce. Thus, an epidemiologic survey was designed to update information on T. gondii infection in pigs from intensive production. Fattening pigs and sows from conventional farms were sampled. Sera were tested with a commercial ELISA for anti-T. gondii antibodies, whereas molecular analysis by 529 bp repetitive element PCR and B1 real-time PCR with subsequent genotyping was performed on heart samples. Statistical analysis was carried out to detect farm management features and sanitary procedures enhancing the risk of infection. At the farm level, 63.6% (7/11) of farms housing sows and 6.7% (1/15) housing fattening pigs scored positive, with individual prevalences of 8.6% (13/151) in sows and 0.5% (1/219) in fattening pigs. T. gondii DNA was detected in eight sows and one fattening pig, and sequencing revealed the presence of all three genotypes (types I, II, and III). Furthermore, the decrease in the biosecurity level enhanced the risk of infection within a farm. The present survey outcomes confirm the spread of T. gondii infection in pig farms in an area of intensive swine production. The application of stricter sanitary procedures may contribute to a further reduction of infection

Seroprevalence of Tick-Borne Infections in Horses from Northern Italy

Tick-borne diseases in horses are considered an emergent problem worldwide; the geographical redistribution of ticks, due to climatic and ecological changes, and the movements of infected horses between different nations play important roles in the spread of tick-borne diseases affecting these hosts. In this study, a survey was planned to estimate the seroprevalence of the Gram-negative bacterium Anaplasma phagocytophilum and the piroplasmid protozoa Babesia caballi and Theileria equi in Italian horses, as well as to evaluate possible risk factors associated with seropositive cases. Serum samples from 261 horses reared in northern Italy were collected and analyzed by indirect immunofluorescence antibody test for the detection of A. phagocytophilum-, B. caballi- and T. equi-specific antibodies. The overall seroprevalence to at least one of the investigated pathogens was 51%; sixty-one horses were seropositive to A. phagocytophilum (23.4%), forty-nine to B. caballi and the same number to T. equi (18.8% each). Seropositivity for more than one of the investigated agents was detected in thirty-two horses and the most common co-infection was observed between B. caballi and T. equi (5.7%). A significant risk factor for all the three pathogens was the elevation above sea level; indeed, the risk of infection was higher with an increase and decrease in elevation for A. phagocytophilum and for B. caballi and T. equi, respectively. Tick control in horses is strongly recommended considering the high seroprevalence values of transmitted pathogens

Lactation Characteristics in Alpine and Nera di Verzasca Goats in Northern Italy: A Statistical Bayesian Approach

In the last three decades, we assisted in a shift in the dairy product preferences in developed countries, with an increase in goat-derived products consumption. As already seen in the bovine sector, the diffusion of goat milk consumption has led to an abandonment of the local breeds in favor of the more productive cosmopolitan breeds, with an intense reduction in the livestock biodiversity. However, in several studies, it has been shown that local breeds present unique features such as a robust behavior towards the local climate and a high quality of their dairy products. This study aims to assess the differences between the basic lactation variables (milk yield, fat %, protein %, lactose % and somatic cell count) in the Alpine goat and the local breed Verzasca goat from a Bayesian point of observation. Results indicate that the Nera di Verzasca goat has overall lactation characteristics similar toa cosmopolite breed such as the Alpine goat, except for the milk daily yield reduced with respect to Alpine goats (1.68 ± 0.84 and 2.14 ± 1.03, respectively). Moreover, the Verzasca goat showed a tendency to maintain a lower level of somatic cell count along with lactation (798 ± 1734 and 1216 ± 2828, respectively). These findings help to increase the knowledge about the Italian local goat breeds and are important in the optic of biodiversity conservation

Comparison of Female Verzaschese and Camosciata delle Alpi Goats' Hematological Parameters in The Context of Adaptation to Local Environmental Conditions in Semi-Extensive Systems in Italy

Simple Summary Livestock biodiversity is being lost at an alarming rate. It is mainly due to genetic selection for desirable traits and the standardization of productions, but this has determined a dramatic reduction in intra- and inter-breed genetic variability. Autochthonous breeds represent a pivotal genetic resource thanks to their environment-specific adaptive features, especially related to rusticity, frugality, fertility, and longevity. In this context, it is of fundamental importance to deepen the knowledge about these breeds' physiology and to take care of their health in the optic of conservation strategies. This study established hematological reference values in female goats of a cosmopolitan (i.e., Camosciata delle Alpi) and an Italian autochthonous (i.e., Verzaschese) breed reared in the same conditions. The influence of breed, age, and season on the hematological parameters was also investigated. The results have shown that variation due to breed, age, and season in blood parameters could be attributed both to physiological changes, such as the ones given by ageing and melatonin effect, and to adaptive genetic processes, for example, towards gastrointestinal parasitism. These findings can be helpful to provide evidence of the importance of recovering endangered/rare local breeds, which are a fundamental heritage for agricultural biodiversity, and local culture. Local livestock breeds are pivotal to ensure sustainable agriculture and represent a real genetic resource in the context of genetic variability reduction. This study aimed at broadening the knowledge about hematological values in female Verzaschese and Camosciata delle Alpi goats (an Italian local and a cosmopolitan goat breed, respectively) and investigating factors affecting them (i.e., breed, age, season). Blood samples were collected monthly from 34 Verzaschese and 37 Camosciata delle Alpi female goats kept under the same semi-extensive farming system for a whole year. The main hematological parameters were evaluated, and descriptive as well as inferential statistical analyses were performed. Reference intervals for complete blood cell count were established and several inter-breed differences were found. In particular, most of the red blood cell parameters were higher in Verzaschese than Camosciata delle Alpi (p < 0.05) suggesting a greater gastrointestinal parasites' resilience of the local breed. The age effect (p < 0.05) was consistent with the literature while the season effect (p < 0.05) could be explained by the melatonin immunostimulant action and gastrointestinal parasitism influence. Overall, differences in blood values could be attributed to physiological changes and adaptive strategies developed through centuries highlighting the remarkable rusticity and adaptation to the environment and farming system of the local breed

Comparison of Female Verzaschese and Camosciata delle Alpi Goats’ Hematological Parameters in The Context of Adaptation to Local Environmental Conditions in Semi-Extensive Systems in Italy

Local livestock breeds are pivotal to ensure sustainable agriculture and represent a real genetic resource in the context of genetic variability reduction. This study aimed at broadening the knowledge about hematological values in female Verzaschese and Camosciata delle Alpi goats (an Italian local and a cosmopolitan goat breed, respectively) and investigating factors affecting them (i.e., breed, age, season). Blood samples were collected monthly from 34 Verzaschese and 37 Camosciata delle Alpi female goats kept under the same semi-extensive farming system for a whole year. The main hematological parameters were evaluated, and descriptive as well as inferential statistical analyses were performed. Reference intervals for complete blood cell count were established and several inter-breed differences were found. In particular, most of the red blood cell parameters were higher in Verzaschese than Camosciata delle Alpi (p < 0.05) suggesting a greater gastrointestinal parasites’ resilience of the local breed. The age effect (p < 0.05) was consistent with the literature while the season effect (p < 0.05) could be explained by the melatonin immunostimulant action and gastrointestinal parasitism influence. Overall, differences in blood values could be attributed to physiological changes and adaptive strategies developed through centuries highlighting the remarkable rusticity and adaptation to the environment and farming system of the local breed