Efficacy of moxidectin injectable and pour-on formulations in a pilot control program against bovine hypodermosis in Southern Italy

Bovine hypodermosis is a myiasis caused by Hypoderma bovis and Hypoderma lineatum (Diptera, Oestridae) larvae, which has a severe economic impact on the livestock industry. Though myiasis is widespread throughout Italy, no nationwide eradication program has ever been planned, unlike in other European Countries. With a view to setting up a national control program, a pilot study was carried out in Southern Italy on 9939 cattle bred in an area with a high prevalence of cattle hypodermosis, using moxidectin 0.5% pour-on (Cydectin, Fort Dodge) and 1% injectable (Cydectin, Fort Dodge) formulations. At the recommended dosage, moxidectin displayed efficacy levels of 99.9% in the pour-on and 100% in the injectable formulation, whereas the microdose (1 mg per head regardless of body weight) was less effective (65.7%). This trial contributed to a significant reduction in infestation rates in the study area and represented the first step through which a national program for eradicating warble fly infestation in Italy.     

Recombinant K39 dipstick immunochromatographic test: a new tool for the serodiagnosis of canine leishmaniasis.

The spread of human leishmaniasis has prompted the scientific community to study dogs as reservoirs for Leishmania infantum. Canine leishmaniasis (CanL) is widespread in the Mediterranean area with a prevalence of up to 50%. The first step toward controlling the disease is to monitor its distribution, mainly in stray dogs. The validity of a recombinant K39 (rK39) dipstick test, commercially available for the serodiagnosis of human leishmaniasis, was evaluated using sera from 165 dogs selected on the basis of positive or negative lymph node smears at parasitological examination. The results were compared with the indirect fluorescent antibody test (IFAT) (cutoff 1:80). Sera from a group of dogs with other diagnosed diseases but negative for leishmaniasis were also tested to evaluate any cross-reactivity. Various procedures were used for testing whole blood samples. The relative specificity of the rK39 dipstick and IFAT was 100% (97 of 97) and 98.97% (96 of 97), whereas the relative sensitivity was 97.06% (66 of 68) and 98.53% (67 of 68), respectively. The results of the dipstick and IFAT corresponded except for 2 sera (k = 0.987). This data confirm the usefulness of rK39 antigen for diagnosing CanL both in symptomatic and asymptomatic dogs. The rK39 dipstick proved to be a rapid, sensitive, and specific test that may be very useful in the field for large-scale screening and also in veterinary practice, requiring minimal equipment and operator expertise.     

Diagnosis and risk factors of Aelurostrongylus abstrusus (Nematoda, Strongylida) infection in cats from Italy

Aelurostrongylus abstrusus (Nematoda, Strongylida) in cats and associated risk factors have been investigated by classical coprology in central (site A) and southern (site B) Italy, i.e. by floatations with sugar and zinc sulphate solutions and a Baermann technique. Overall, 227 individual stool samples (i.e. 162 from site A and 65 from site B) were processed and risk factors were related to infection by binary logistic multiple-regression models. The presence of A. abstrusus was used as the dependent variable and the epidemiological data as independent variables. Twenty-eight (17.3%) and 12 (18.5%) cats were positive for A. abstrusus in sites A and B, respectively, both using the Baermann and flotation with zinc sulphate solution methods. Six and three (from site A and B, respectively) of the infected cats were negative for A. abstrusus when faeces were processed with sugar solution. Stray and free-ranging (P=0.357), young cats (P=0.008) and the presence of respiratory symptoms (P=0.000) were risk factors for A. abstrusus infection. The results suggest that A. abstrusus is spread in Italy. Furthermore, these results possibly include infection in the differential diagnosis of feline respiratory diseases and offer a correct diagnostic approach by using either a Baermann method or flotation with zinc sulphate solution in suspected cases.     

A comparison of coprological, serological and molecular methods for the diagnosis of horse infection with Anoplocephala perfoliata (Cestoda, Cyclophyllidea)

Anoplocephala perfoliata (Cestoda, Cyclophyllidea), the commonest intestinal tapeworm of horses, can cause colic, intussusceptions, ileal impactions and intestinal perforations. Common diagnostic techniques for A. perfoliata infection, i.e. coprology and serology, show inherent limitations in terms of sensitivity and specificity and new approaches are thus required. Hence, the present study compared the reliability of coprological, serological (i.e. ELISA) and molecular (i.e. nested PCR) methods in detecting A. perfoliata infection in naturally infected horses and in horses treated with a combination of ivermectin and praziquantel. Of 42 horses subjected to coprological examination, 16 and 26 resulted negative and positive, respectively for the presence of A. perfoliata eggs at the coprological examination. The 26 coprologically positive animals were also positive by nested PCR. Fifteen out of the 16 horses coprologically negative were negative at the molecular assay, while one yielded a PCR product detectable on an agarose gel. Eighteen out of 26 positive horses were treated with a combination of ivermectin 18.7 mg/g and praziquantel 140.3mg/g and resulted subsequently negative by coprology and nested PCR performed 2 weeks after treatment. All infected and untreated animals had a high ELISA test optical density indicating high infection intensity and associated risk of colic. However, high optical density values were also obtained in four horses post-treatment and in three horses that were negative on molecular and coprological analysis. The results of the present work indicate that the nested PCR assay represents a valid method for the specific molecular detection of A. perfoliata in faecal samples collected from naturally infected horses and may have advantages over coprological and serological approaches for diagnosing A. perfoliata infection.     

Molecular diagnosis of equid summer sores

Equine cutaneous habronemosis, also known as "summer sores", is a parasitic infection caused by larvae of Habronema microstoma and Habronema muscae (Nematoda, Spirurida) released by dung-inhabiting fly vectors on abraded skin, skin wounds or muco-cutaneous transition sites. Larvae induce a local inflammatory reaction characterised by itching, granulomatous, ulcerated and, often non-healing, lesions. The diagnosis of summer sores may be unreliable mainly because of the limits of clinical and microscopic examination. The applicability of a semi-nested PCR assay developed for the diagnosis of gastric habronemosis has been herein demonstrated for the detection of the cutaneous infection. The potential applicability of this diagnostic tool may have for clinical and epidemiological studies of cutaneous habronemosis in equids is discussed.     

Occurrence of anthelmintic resistant equine cyathostome populations in central and southern Italy

In the present survey, 276 horses bred on 16 farms located in central and southern Italy were investigated for the presence of drug resistant cyathostomes by a Fecal Egg Count Reduction Test (FECRT). Sixteen to 20 animals were selected on each farm and randomly assigned to one of four equally sized treatment groups. Groups were treated with fenbendazole, pyrantel pamoate, ivermectin or moxidectin. Resistance to fenbendazole was declared on six farms (37.5%) and suspected in two farms (12.5%), with FECR values ranging from 41% to 88.3%. Resistance to pyrantel was found in two farms (12.5%) and was suspected in one case (6.2%), with FECR values ranging from 43% to 85.4%. Macrocyclic lactones remained effective on all farms. Only cyathostome third stage larvae (L3) were found in fecal cultures after treatment. This paper reports the first wide survey conducted in Italy for anthelmintic resistance in equine cyathostomes. The results indicate that multiple drug resistant equine cyathostomes are present in the central and southern regions of Italy. These data call for a geographically and numerically broader investigation of horse farms in all regions and for the development and implementation -among veterinarians, owners and managers of a plan to reduce the expansion of these anthelmintic resistant populations and control these important parasites.     

Fermentation Technologies for the Optimization of Marine Microbial Exopolysaccharide Production

In the last decades, research has focused on the capabilities of microbes to secrete exopolysaccharides (EPS), because these polymers differ from the commercial ones derived essentially from plants or algae in their numerous valuable qualities. These biopolymers have emerged as new polymeric materials with novel and unique physical characteristics that have found extensive applications. In marine microorganisms the produced EPS provide an instrument to survive in adverse conditions: They are found to envelope the cells by allowing the entrapment of nutrients or the adhesion to solid substrates. Even if the processes of synthesis and release of exopolysaccharides request high-energy investments for the bacterium, these biopolymers permit resistance under extreme environmental conditions. Marine bacteria like Bacillus, Halomonas, Planococcus, Enterobacter, Alteromonas, Pseudoalteromonas, Vibrio, Rhodococcus, Zoogloea but also Archaea as Haloferax and Thermococcus are here described as EPS producers underlining biopolymer hyperproduction, related fermentation strategies including the effects of the chemical composition of the media, the physical parameters of the growth conditions and the genetic and predicted experimental design tools.     

Prediction of dry matter, fat, pH, vitamins, minerals, carotenoids, total antioxidant capacity, and color in fresh and freeze-dried cheeses by visible-near-infrared reflectance spectroscopy

Visible-near-infrared reflectance spectroscopy was used to predict dry matter, fat, pH, retinol, alpha-tocopherol, beta-carotene, xanthophylls, sodium chloride, calcium, potassium, magnesium, zinc, total antioxidant capacity, brightness, redness, and yellowness in both fresh and freeze-dried cheeses. A total of 445 cheeses of four cheese varieties were investigated. Composition of samples was analyzed by reference methods. Samples were scanned (400-2500 nm) and predictive equations were developed using modified partial least-squares with both cross-validation and external validation. Coefficient of determination (R(2)) and residual predictive deviation (RPD) in external validation were satisfactory for dry matter (0.97; 5.99), fat (0.90; 3.22), beta-carotene (0.92; 3.43), sodium chloride (0.89; 2.94), calcium (0.95; 4.62), Zinc (0.93; 3.75), brightness (0.96; 4.88), redness (0.96; 5.23), and yellowness (0.93; 3.73) in fresh cheeses. Poor predictions were obtained for pH, retinol, alpha-tocopherol, xanthophylls, potassium, magnesium, and total antioxidant capacity (R(2) < 0.81; RPD < 3).     

Effect of the pH of heating on the qualitative and quantitative compositions of the sera of reconstituted skim milks and on the mechanisms of formation of soluble aggregates

The effect of the pH of heating (6.3-7.3) on the composition of sera in reconstituted skimmed milks was investigated. A combination of SDS-PAGE analysis and size exclusion chromatography (SEC) combined with an original approach to the analysis of the SEC profiles was performed. The composition of the sera varied greatly when the pH of heating was adjusted below and above the natural pH of milk. The formation, composition, and concentration of heat-induced soluble complexes depended on the combination of the effect of adjusting the pH of the milk and the heat treatment. Two types of mechanism for the formation of soluble aggregates appeared to exist, depending on the pH of the milk. The first type results from the formation of WP/kappa-casein aggregates at the surface of the micelle, and these were detached partially into the serum in larger amount as the pH increased up to 6.7, where it reaches a maximum. The second type of complexes, whose amount increased as the pH of heating increased from 6.7 to 7.3, may be formed between caseins (kappa- but also perhaps some alpha(s)-casein) and aggregated WP resulting in complexes that are smaller in size and with a higher kappa-casein/whey protein ratio than the first type.     

Association of SNP variants of MHC Class II DRB gene with thermo-physiological traits in tropical goats

Host defense in vertebrates depend on many secreted regulatory proteins such as major histocompatibility complex (MHC) class II which provide important regulatory and effector functions of T cells. Gene polymorphism in the second exon of Capra-DRB gene in three major Nigerian goat breeds [West African Dwarf (WAD), Red Sokoto (RS), and Sahel (SH)] was analyzed by restriction fragment length polymorphisms (RFLP). Four restriction enzymes, BsaHI, AluI, HaeIII, and SacII, were utilized. The association between the polymorphic sites and some heat tolerance traits were also investigated in a total of 70 WAD, 90 RS, and 50 SH goats. Fourteen different types of alleles identified in the Nigerian goats, four of which were found in the peptide coding region (A57G, Q89R, G104D, and T112I), indicate a high degree of polymorphism at the DRB locus in this species. An obvious excess (P?<?0.01) of non-synonymous substitutions than synonymous (dN/dS) in this locus is a reflection of adaptive evolution and positive selection. The phylogenetic trees revealed largely species-wise clustering in DRB gene. BsaHI, AluI, HaeIII, and SacII genotype frequencies were in Hardy-Weinberg equilibrium (P?>?0.05), except AluI in RS goats and HaeIII in WAD goats (P?<?0.05). The expected heterozygosity (H), which is a measure of gene diversity in the goat populations, ranged from 0.16 to 0.50. Genotypes AA (BsaHI), GG, GC and CC (AluI) and GG, GA, AA (HaeIII) appeared better in terms of heat tolerance. The heat-tolerant ability of SH and RS goats to the hot and humid tropical environment of Nigeria seemed better than that of the WAD goats. Sex effect (P?<?0.05) was mainly on pulse rate and heat stress index, while there were varying interaction effects on heat tolerance. Variation at the DRB locus may prove to be important in possible selection and breeding for genetic resistance to heat stress in the tropics.