Liquid chromatography-tandem mass spectrometric ion-switching determination of chlorantraniliprole and flubendiamide in fruits and vegetables

The anthranilic and phthalic diamides, chlorantraniliprole (CAP) and flubendiamide (FLU), respectively, represent a new class of very effective insecticides that activate the ryanodine-sensitive intracellular calcium release channel (ryanodine receptor). This paper reports an analytical method for the simultaneous determination of the two insecticides on fruits and vegetables by liquid chromatography-electrospray tandem mass spectrometry operated in the positive and negative ionization switching mode. The two diamides were extracted with acetonitrile and separated on a Zorbax Column Eclipse XDB C8 (4.6 mm x 150 mm i.d., 3 microm) by isocratic elution with a mobile phase consisting of acetonitrile and water with 0.1% formic acid pumped at a flow rate of 0.4 mL/min. The diamides were selectively detected by multiple reaction monitoring for transitions of proton adduct precursor ions simultaneously: positive m/z 484.3-->285 for CAP, m/z 445.5-->169 for internal standard, and negative m/z 681.4-->253 for FLU. For CAP calibration in the positive mode was linear over a working range of 2 to 1000 microg/L with r > 0.992. The limit of detection (LOD) and limit of quantification (LOQ) for CAP were 0.8 and 1.6 microg/kg, respectively. For FLU in the negative mode the corresponding values were 1-1000 microg/L for linear working range, with r > 0.996 and 0.4 and 0.8 microg/L for LOD and LOQ, respectively. Moreover, the presence of interfering compounds in the fruit and vegetable extracts was found to be minimal. Due to the linear behavior of the MS detector response for the two analytes, it was concluded that the multiple reaction transitions of molecular ions in the ion-switching mode can be used for analytical purposes, that is, for identification and quantification of diamides in fruit and vegetable extracts at trace levels.     

Clinical evaluation and microbiota analysis in 9 dogs with antibiotic‐responsive enteropathy: A prospective comparison study

BackgroundAntibiotic‐responsive enteropathy (ARE) is diagnosed by excluding other causes of diarrhea and when there is a short‐term response to administration of antibiotics.ObjectivesTo characterize the gut microbiota and clinical trend of dogs with suspected ARE and to evaluate the variation in microbiota before (T0), after 30 days (T30) of tylosin treatment, and 30 days after discontinuation of treatment (T60). A further objective was to evaluate whether changes in gut microbiota are related to relapses of diarrhea when the therapy is tapered.AnimalsStudy sample (group A) was composed of 15 dogs with chronic diarrhea, group B was composed of 15 healthy dogs. Group A was given tylosin for 30 days.MethodsA multicentric prospective study. Clinical Indexes, fecal score, and samples for microbiota analysis were collected at T0, T30, and T60 in group A and T0 and T30 in group B. The gut microbiota was analyzed via 16S ribosomal RNA gene. Qiime2 version 2020.2 was used to perform bioinformatic analyses, and Alpha‐ and Beta‐diversity were computed.ResultsDiarrhea recurred after T30 in 9 of 14 dogs, which were classified as affected by ARE. At T0, a difference was noted in the beta‐diversity between groups (Bray Curtis metric P = .006). A T0‐T30 difference in alpha‐diversity was noted in group A (Shannon index P = .001, Faith PD P = .007).Conclusions and Clinical ImportanceAlthough tylosin influences the microbiota of dogs with ARE, we failed to find any specific characteristic in the microbiota of dogs with ARE.     

Sartorius muscle contracture in a German shepherd dog

Objective— To report sartorius muscle contracture in a dog.
Study Design— Case report.
Animals— A 5-year-old, male, German Shepherd dog.
Methods— After a 2-month history of right hindlimb lameness was treated by tenectomy of the caudal sartorius muscle for sartorius muscle contracture.
Results— One month after surgery there was complete remission of clinical signs; however, after 12 months, there was partial reunion of sartorius caudal belly and onset of gracilis muscle contracture.
Conclusions— Contracture of the sartorius muscle in dogs is seemingly rare and similar to gracilis muscle contracture with which it may be confused. Differentiation requires knowledge of muscular anatomy, careful palpation, and surgical inspection.
Clinical Relevance— Sartorius muscle contracture can be treated by tenectomy or removal of contracted muscle tissue, however, long-term outcome suggests the need to develop approaches that prevent recurrence caused by scarring of transected tissues.     

Aggressive dogs are characterized by low omega-3 polyunsaturated fatty acid status

Canine aggressive behaviour is one of the most common problems being reported by dog owners. However, the biochemical basis of this phenomenon remains unclear. In humans, alterations in omega-3 plasma polyunsatured fatty acids and elevated omega6/omega-3 ratio have been linked to behavioural alterations, including aggression. Thus far, however, the relationship between plasma polyunsatured fatty acid status and aggression has not been investigated in the dog. In the present study we sought to investigate whether polyunsatured fatty acid status could be altered in plasma of pathologically aggressive Canis familiaris. Eighteen adult male German Shepherd dogs, aged 4.9 ± 0.9 years, showing no clinical signs but aggression, were investigated. Eighteen healthy male dogs, aged 4.8 ± 0.7 years, with a negative history of behavioural and neurological disorders served as controls. Baseline fasting plasma polyunsatured fatty acid composition was determined by gas chromatography. Compared to normal dogs, aggressive dogs showed lower docosahexaenoic acid (22:6 n-3) concentrations and a higher omega6/omega-3 ratio. In addition, they showed reduced cholesterol and bilirubin concentrations compared to their normally behaving counterparts. Altogether, our results suggest that low omega-3 fatty acids may adversely impact behaviour in dogs, resulting in greater propensity to aggression. However, given the cross-sectional design of our study, we cannot claim any causal relationship between the presence of alterations in fatty acid status and canine aggressiveness. Whether omega-3 fatty acids supplementation may be useful to reduce aggressive behaviour in the dog deserves further investigation.     

Fungal Flora of Normal Eyes in Healthy Newborn Foals Living in the Same Stud Farm in Italy

The aim of this study was to assess the presence of fungal flora in conjunctiva of newborn foals revealed by means of repeated culture examinations and to compare prevalences in foals and their mares. Fifty-four healthy foals and their mothers were used. Ocular samples were collected from foals immediately after parturition (D0), and at days 2 (D2), 7 (D7), 14 (D14), and 28 (D28), and at D0 from mares. Samples were seeded onto Sabouroud dextrose agar and malt extract agar, incubated at 25°C and examined daily over a 10-day period. Twenty-two of 54 foals were positive for conjunctival fungi at D0, 13 of 54 at D2, 16 of 54 at D7, 19 of 54 at D14, and 13 of 54 at D28. The most frequently isolated fungi were Penicillium species and Aspergillus versicolor. Colony-forming units ranged from 1 to 5. One of 54 foals yielded the same fungus more than two times consecutively; 23 of 54 mares were positive for conjunctival fungi, and the most frequently recovered were Penicillium species, Aspergillus species, and Saccharomyces cerevisiae. Significant differences among prevalences, both at different sampling times in foals, and between mares and foals at D0, were not observed. A fungal contamination of conjunctival fornix in foals at birth and during their first month of life was reported. Fungal isolation in foals at D0 might be explained by the presence of fungal contamination within mares' genitalia, whereas positive cultures during the first month of life probably represent transient seeding from the environment. Only once was the same fungus isolated from the same foal more than two times consecutively.     

An Ibero-American inter-laboratory trial to evaluate serological tests for the detection of anti-<Emphasis Type="Italic">Neospora caninum</Emphasis> antibodies in cattle

We carried out an inter-laboratory trial to compare the serological tests commonly used for the detection of specific Neospora caninum antibodies in cattle in Ibero-American countries. A total of eight laboratories participated from the following countries: Argentina (n = 4), Brazil (n = 1), Peru (n = 1), Mexico (n = 1), and Spain (n = 1). A blind panel of well-characterized cattle sera (n = 143) and sera representative of the target population (n = 351) was tested by seven in-house indirect fluorescent antibody tests (IFATs 1–7) and three enzyme-linked immunosorbent assays (ELISAs 1–3; two in-house and one commercial). Diagnostic performance of the serological tests was calculated and compared according to the following criteria: (1) the “Pre-test information,” which uses previous epidemiological and serological data; (2) the “Majority of tests,” which classifies a serum as positive or negative according to the results obtained by most tests evaluated. Unexpectedly, six tests showed either sensitivity (Se) or specificity (Sp) values lower than 90%. In contrast, the best tests in terms of Se, Sp, and area under the ROC curve (AUC) values were IFAT 1 and optimized ELISA 1 and ELISA 2. We evaluated a high number of IFATs, which are the most widely used tests in Ibero-America. The significant discordances observed among the tests regardless of the criteria employed hinder control programs and urge the use of a common test or with similar performances to either the optimized IFAT 1 and ELISAs 1 and 2.     

ACVIM consensus statement on the diagnosis of immune‐mediated hemolytic anemia in dogs and cats

Immune‐mediated hemolytic anemia (IMHA) is an important cause of morbidity and mortality in dogs. IMHA also occurs in cats, although less commonly. IMHA is considered secondary when it can be attributed to an underlying disease, and as primary (idiopathic) if no cause is found. Eliminating diseases that cause IMHA may attenuate or stop immune‐mediated erythrocyte destruction, and adverse consequences of long‐term immunosuppressive treatment can be avoided. Infections, cancer, drugs, vaccines, and inflammatory processes may be underlying causes of IMHA. Evidence for these comorbidities has not been systematically evaluated, rendering evidence‐based decisions difficult. We identified and extracted data from studies published in the veterinary literature and developed a novel tool for evaluation of evidence quality, using it to assess study design, diagnostic criteria for IMHA, comorbidities, and causality. Succinct evidence summary statements were written, along with screening recommendations. Statements were refined by conducting 3 iterations of Delphi review with panel and task force members. Commentary was solicited from several professional bodies to maximize clinical applicability before the recommendations were submitted. The resulting document is intended to provide clinical guidelines for diagnosis of, and underlying disease screening for, IMHA in dogs and cats. These should be implemented with consideration of animal, owner, and geographical factors.     

Outcome and failure patterns of localized sinonasal lymphoma in cats treated with first‐line single‐modality radiation therapy: A retrospective study

Failure rate and site are not well defined in localized sinonasal lymphoma in cats treated with radiotherapy. In this study, we describe (a) failure pattern, (b) outcome, (c) influence of previously reported prognostic variables on the outcome in cats with suspected localized sinonasal lymphoma. In this multi‐institutional retrospective study, we included 51 cats treated with single‐modality radiotherapy. Cats were irradiated using 10x4.2Gy (n = 32), 12x3Gy (n = 11) or 5x6Gy (n = 8). Regional lymph nodes were prophylactically irradiated in 24/51 cats (47.1%). Twenty‐five cats (49.0%) developed progressive disease: progression was local (nasal) in five (9.8%), locoregional (nodal) in two (3.9%), local and locoregional in three (5.9%), systemic in nine (17.6%) and both local and systemic in six cats (11.8%). No cat receiving prophylactic nodal irradiation had progression in the locoregional lymph nodes. The median time to progression was 974 days (95%CI: 283;1666), with 58% and 53% of cats free of progression at 1 and 2 years, respectively. Median overall survival was 922 days (95%CI: 66;1779) with 61% and 49% alive at 1 and 2 years, respectively. Half of the cats that died of relapse/progression (13/26) died within 6 months of treatment, suggesting possible shortcomings of staging, rapid dissemination of disease or sequential lymphomagenesis. None of the prognostic factors evaluated were predictive of outcome (prednisolone use, anaemia, nasopharyngeal involvement, modified canine Adams tumour stage, protocol, total dose). Radiotherapy is an effective treatment for localized sinonasal lymphoma with a long time to progression. However, in one‐third of the cats, systemic disease progression occurs soon after radiotherapy.     

Mycotoxigenic fungi contaminating greenhouse-grown tomato fruit and their alternative control

European Journal of Plant Pathology - Tomato fruit represent a valuable but perishable commodity due to postharvest fungal spoilage. In the present investigation, greenhouse-grown tomatoes were...