Gastrointestinal parasites, liver flukes and lungworms in domestic ruminants from central Italy

Introduction - In ruminants, gastrointestinal, liver and lung parasites may represent a limiting factor for farm production. Aim - The aim of this study was to evaluate the occurrence of gastrointestinal, liver and lung parasites in adult ruminants living in two different areas of Tuscany, central Italy. Materials and methods - Between April 2012 and December 2014, 178 adult ruminants (76 cattle, 61 sheep and 41 goats) from 16 extensive farms located in two different areas (A1 and A2) of Tuscany, were examined to assess the occurrence of gastrointestinal parasites, liver flukes and lungworms. A1 included 111 animals from farms located in flat areas subject to water stagnation in rainy seasons, while A2 included 67 animals from farms located in hilly and drier areas. Individual faecal samples collected from all animals were analysed using qualitative and quantitative parasitological techniques. A total of 94 animals were examined for Fasciola hepatica also by using two commercial Elisa kits for the detection of faecal antigens and antibodies in serum, respectively. Data were statistically analysed. Results and discussion - An overall prevalence of 83.7% was found in the examined animals. Higher prevalence values (p<0.001) were found in small ruminants than in cattle and in Area 2 compared to Area 1. With regard to isolated parasites, gastrointestinal strongyles and coccidia were prevalent in all ruminant species and in both areas, while the prevalence of F. hepatica was higher in small ruminants and in Area 1 than in cattle and Area 2, respectively. Conclusion - Results indicated that in both areas and in all ruminant species, gastrointestinal parasites and liver flukes require more effective control measures

Incidental Detection of Onchocerca Microfilariae in Donkeys (Equus asinus) in Italy: Report of Four Cases

This paper reports the occurrence for the first time in Italy of autochthonous Onchocerca infection in donkeys. Four jennies, bred on the same farm, were referred to the Veterinary Teaching Hospital of Pisa for a check-up on ovarian activity (n = 3) or for veterinary support during the delivery (n = 1). Microfilariae were incidentally detected during the blood smear examination of one jenny. Peripheral blood samples were then collected from the other three jennies and the presence of microfilariae was investigated by Knott's test. Circulating unsheathed microfilariae were identified in all the animals. The level of microfilaraemia was between 1 and 31 microfilariae in 2 mL of blood. Hematological changes showed moderate eosinophilia in one case or both remarkable eosinophilia and basophilia in another case. Based on molecular findings by PCR and sequencing, the microfilariae showed 98% sequence similarity with Onchocerca sp. in the NCBI GenBank database (Accession No.: MK541848.1). The present report provides evidence that Onchocerca is an etiological agent of parasitic infection in donkeys in Italy. Our findings highlight the importance of screening in donkeys for Onchocerca even in the absence of clinical indications

Preliminary validation study of Paraoxonase-1 in horses

Paraoxonase-1 (PON-1) is an anti-oxidant enzyme associated with high-density lipoproteins in blood. PON-1 is a negative acute-phase protein being its plasmatic activity reduced during inflammation due to consumption by oxidants. Considering the possible clinical usefulness of PON-1 as an early inflammatory marker this is a preliminary validation study in horses. Serum PON-1 activity was measured in 69 clinically healthy animals (31 adult female, 18 geldings, 11 stallions, 9 foals) using an enzymatic method adapted from other species. In order to preliminarily assess the possible utility of PON-1 as a marker of Systemic Inflammatory Response Syndrome (SIRS), blood from 6 sick foals, classified according to a validated SIRS scale, was analyzed. Intra- and inter-assay imprecision were assessed by repeated analysis of pooled samples and evaluation of coefficient of variations (CV). Accuracy was indirectly evaluated through linearity under dilution (LUD) and spiking recovery test (SRT). Results of the different groups of healthy horses were compared to each other with a Friedmann test with Bonferroni correction. The method is precise (inter- and inter-assay CVs <5%) and accurate (LUD and SRT fit the linear model). PON-1 activity was higher in foals and in adult females (mean ± SD: 63.7±15.5 and 60.8±10.1, respectively) than in geldings and adult males (52.5±10.2 and 47.2±7.7, respectively). In 3/6 SIRS foals PON-1 activity was lower than the lowest percentile of distribution of healthy foals. This study demonstrated that the method of measurement of PON-1 activity in horses is precise and accurate and PON-1 may be a marker of SIRS

Using roquefortine C as a biomarker for penitrem A intoxication in a beef herd

Fifteen grazing beef cattle and calves presented a history of neurological signs like ataxia, intentional head tremors, muscle twitching. Nervous ketosis, nervous BVD, BHV-1,5, tremorgenic intoxication from hay, and Listeriosis were considered as differential diagnosis. Blood samples were collected. Inspection of hay bales showed large white dusty and moldy areas. Samples were taken and analyzed. Altered hay was immediately removed in all animals’ stock. No alterations were found in blood tests. Food analysis showed high concentrations of Roquefortine C (RC) (345 μg/kg DM). Tremorgenic syndrome has been reported in Penitrem A (PA) intoxication, but PA is difficult to isolate in laboratory conditions. Both RC and PA are produced by Penicillum spp. RC has been associated with PA in tremorgenic toxicosis in dogs and it might be considered a valuable diagnostic marker for PA intoxication. The neurological signs were due to tremorgenic intoxication after feeding of spoiled forage contaminated with mycotoxines

evaluation of three commercial rapid kits to detect cryptosporidium parvum in diarrhoeic calf stool

The aim of this study was to evaluate three commercially available rapid immunochromatographic tests for detection of Cryptosporidium parvum antigens in faeces of naturally infected neonatal diarrhoeic calves. FASTest® CRYPTO strip, FASTest® CRYPTO-GIARDIA Strip and TETRASTRIPS® were compared for their sensitivity, specificity, positive predictive value and negative predictive value using a cumulative positivity as gold standard. In addition, the agreement between each test and the gold standard was evaluated by Cohen's Kappa (k) value. The highest infection rate was observed by FASTest® CRYPTO-GIARDIA Strip (65.15%), followed by FASTest® CRYPTO strip (63.64%) and TETRASTRIPS® (56.06%,). A very good diagnostic performance of all the three tests was observed. FASTest® CRYPTO strip (k = 0.935) and FASTest® CRYPTO-GIARDIA Strip (k = 0.968) had the highest sensitivity (100%) while TETRASTRIPS® (k = 0.875) had the highest specificity (100%). Eimeria spp oocysts were present in six samples but cross-reaction with this protozoan was not observed. These assays were not time-consuming and very easy to perform and to read. Based on our results, we recommend the use of FASTest® CRYPTO strip, FASTest® CRYPTO-GIARDIA Strip or/and TETRASTRIPS® for detection of C. parvum antigens in faeces of neonatal diarrhoeic calves

Evaluation of three commercial rapid kits to detect Cryptosporidium parvum in diarrheic calf stool

The aim of this study was to evaluate three commercially available rapid immunochromato-graphic tests for detection of Cryptosporidium parvum antigens in faeces of naturally infected neonatal diarrhoeic calves. FASTest (R) CRYPTO strip, FASTest (R) CRYPTO-GIARDIA Strip and TETRASTRIPS (R) were compared for their sensitivity, specificity, positive predictive value and negative predictive value using a cumulative positivity as gold standard. In addition, the agreement between each test and the gold standard was evaluated by Cohen's Kappa (k) value. The highest infection rate was observed by FASTest (R) CRYPTO GIARDIA Strip (65.15%), followed by FASTest (R) CRYPTO strip (63.64%) and TETRASTRIPS (R) (56.06%,). A very good diagnostic performance of all the three tests was observed. FASTest (R) CRYPTO strip (k= 0.935) and FASTest (R) CRYPTO-GIARDIA Strip (k= 0.968) had the highest sensitivity (100%) while TETRASTRIPS (R) (k= 0.875) had the highest specificity (100%). Eimeria spp oocysts were present in six samples but cross-reaction with this protozoan was not observed. These assays were not time-consuming and very easy to perform and to read. Based on our results, we recommend the use of FA

Bartonella infection in asymptomatic horses and donkeys from Tuscany, Central Italy.

Objective To investigate the occurrence of Bartonella sp. infection in asymptomatic horses and donkeys living in Tuscany, Central Italy. Methods Blood samples were collected from 77 horses and 15 donkeys and tested by indirect immunofluorescent test to detect antibodies against Bartonella sp. and by PCR to detect the pathogen. Results Fifty-four (58.69%; 95% CI: 47.95%–68.87%) animals, 9 donkeys and 45 horses, were seropositive with antibody titers ranging from 1:64 to 1:512. PCR assays detected 9 horses positive for Bartonella sp. and 3 donkeys for Bartonella henselae genotype I. Conclusions The detected sero-prevalence suggests a common and frequent exposure of equids living in Central Italy to bartonellae and PCR results show that Bartonella sp. infection is possible both in horses and donkeys. At the best of our knowledge, this is the first report of Bartonella henselae infection in donkeys

Evaluation of Some Physical, Haemathological and Clinical Chemistry Parameters in Healthy Newborn Italian Holstein Calves

Abstract: The aim of the present study was to investigate some physical, haematological and clinical chemistry parameters in the newborn Italian Holstein calf at birth and at 24 h of life, to evaluate changes during the immediate post-partum period. Forty-six Italian Holstein Friesian calves were included in this study. Heart rate, respiratory rate and body temperature were recorded at birth and at 24 h of life. The time needed to raise the head, acquire sternal recumbency, stand up were also recorded. Blood samples were collected before first feeding and at 24 h of age and CBC count, L-lactate, glucose and total protein concentrations were evaluated. The head was raised immediately in 46/46 calves, suckling reflex was acquired within 12±9 min, sternal position in 5±2 min and newborn stood up in 38±30 min. Some of the physical data, haematological and biochemical values showed statistical differences between birth and 24 h of age. The results from this study provide some information about physical and laboratory data of Italian Holstein Friesian calves, at birth and at 24 h of life. Our results confirm that several clinical and laboratory values in newborn calves differ from adult reference intervals and from calves of different breeds

A possible tremorgenic mycotoxicosis by Roquefortine C in a bovine herd

A total of 15 beef cows and calves were referred for history of neurological signs. The animals (12/15 Chianina breed, 3/15 Limousine) were grazing in 300 ha area, fed with grass and hay. Inspection of the hay reveled macroscopic alterations, consisting of diffuse and heavy mold contamination of many hay bales. Due to the not cooperative attitude, the animals were only visually examined in the field; the neurological signs observed were ataxia, intentional head tremors and muscle twitching. Only 3 calves with severe neurological signs were housed in a medication area and underwent a complete clinical exam. All 3 calves showed intentional head tremors and muscle twitching; 1/3 presented severe ataxia and stiffness gait, while 2/3 calves were recumbent and unable to rise. The most important clinical data were: hyperthermia, tachypnea, tachycardia and long capillary refill time. The neurological examination showed deficits of V and VII cranial nerves. Calves could swallow, but they were unable to grab the food. Based on history and clinical examination the following differential diagnoses were considered: tremorgenic mycotoxicosis, nervous ketosis, nervous BVD form, BHV1-5, Listeriosis and WMD. Blood samples were collected for CBC count and biochemistry panel (TP, urea, creatinine, total and direct bilirubin, GGT, AST, CPK, Mg, Se and vit E), urinalysis was performed for ketone bodies. Calves were also tested for infectious diseases (Listeriosis, BVD, BHV 1-5). Multiple samples of altered hay were analyzed for mycotoxins and hay balls were removed in all animals’ stock. The grazing animals recovered spontaneously within 1 week along with 2/3 hospitalized calves, while 1/3 calf was euthanized due to poor general conditions. CBC, biochemistry panel, vit E and oligo-minerals resulted within normal ranges and no positivity for infectious agents were detected. Food analysis showed high concentrations of roquefortine C (RC): 345 μg/kg DM. Presence of RC in livestock food is highly reported, in particular in visibly moldy areas (1). RC intoxication causes anorexia, paralysis and several reports attribute it neurotoxic properties (2). In mice experimental intoxications induced muscle contractions, ataxia, prostration and intermittent seizures. RC intoxication, resembling penitrem A (PA) intoxication, has been reported in dogs. Moreover, RC is considered a sensitive biomarker for PA exposure. PA is a tremorgenic fungal toxin which intoxication causes ataxia, tachypnea, and sustained tremors. The pathophysiological mechanism by which mycotoxins affect the CNS is unknown but the biochemical lesions are reversible. Diagnosis is based on the clinical signs, demonstration of the mycotoxins in the feed and identification of the fungal elements in blood and feces. Affected animals recover completely when they are removed from infected pastures. Based on neurological signs, recovery after altered food removing and results of food analysis, the diagnosis of tremorgenic intoxication was hypothesized. Limits of this report are: lack of PA dosage in the food and lack of RC and PA evaluation in blood and feces of affected animals

Validation of a Paraoxon-based method for measurement of Paraoxonase (PON-1) activity and establishment of RI in horses

Paraoxonase-1 (PON-1) is an anti-oxidant compound considered as negative acute phase protein in animals (Rossi et al., 2013) and people (Novak et al., 2010). The paraoxon-based method for measurement of PON-1 in equine serum has not yet been validated.The aim of this study is to validate a paraoxon-based method to measure PON-1 and to establish reference intervals (RIs) in healthy horses and foals.120 horses (40 geldings, 40 stallions, 40 mares; median age: 11 years; 57 Warmbloods, 46 Trotters) and 55 foals (27 females, 28 males; median age: 47 days; 22 Warmbloods, 31 Trotters) considered healthy after physical examination and biochemistry were examined. Horses were grouped by breed: Thoroughbreds, Trotters, Warmbloods, Draft horses and Ponies. Serum PON-1 was measured with an automated spectrophotometer and an enzymatic method validated in other species (Giordano et al., 2013). After the analytical validation (precision, accuracy, interference studies), RIs were determined using the Reference Value Advisor software, according to ASCVP guidelines (Friedrichs et al., 2012). The possible gender-, age- and breed-related differences were statistically investigated.The paraoxon-based method was precise (CVs <4.0%) and accurate (P<0.001 in linearity under dilution and spike-recovery testing) but is affected by interference from mild bilirubinemia, severe lipemia or hemoglobinemia. The RIs recorded in the whole population was 38.1-80.8 U/mL. According to the Harris and Boyd test, separate RIs are recommended only for adult females and for Warmblood and Trotter adults (Figure 1).This study demonstrated that analytical performances of the paraoxon-based method for measurement of PON-1 in horses are acceptable. PON-1 is lower in horses than in other species.If future studies will demonstrate that oxidative stress induces a significant decrease of PON-1, this results will be useful to correctly classify healthy and sick horses; PON-1 could be used, as in human medicine, as a marker of oxidative stress