Noninferiority study evaluating the efficacy of a teat disinfectant containing copper and zinc for prevention of naturally occurring intramammary infections in an automatic milking system.

The aim of this study was to demonstrate the noninferiority of a novel teat disinfectant based on copper and zinc (ZkinCu; Copper Andino, Santiago de Chile, Chile) compared with a previously proven glycolic acid active disinfectant (OceanBlu; DeLaval, Kansas City, MO) as a positive control, with respect to the incidence of new intramammary infections under natural challenge conditions on a commercial robotic dairy farm. This study was conducted in 6 robotic pens of approximately 60 milking cows each. The pens were randomly assigned to 1 of the 2 studied disinfectants. Throughout the 8 wk study, the same pre- and post-milking teat disinfectant was used in each pen. The same milking procedures were used in each robot throughout the study. Pre-milking hygiene consisted of applying the disinfectant (OceanBlu or ZkinCu) with the robotic arm. The same product was applied on the teats after milking. At the beginning of the study, all quarters of all study cows were sampled. In successive samplings (wk 2, 4, 6, and 8), composite milk samples were collected on farm to determine SCC. Once composite SCC results were available (2 d) and based on an SCC of ≥100,000 cells/mL, quarter milk samples underwent bacteriological culture. Clinical mastitis was identified by study personnel. Intramammary infection in biweekly quarter milk samples was determined based on composite SCC levels (≥100,000 cells/mL) and the presence of bacteria. A new IMI was defined as a quarter in which the organism isolated was not present in the previous bacteriological sample, or the previous composite SCC sample was <100,000 cells/mL. Clinical mastitis samples were also considered to be new IMI. The trial was designed as a positive control field trial, in which the objective was to show noninferiority of ZkinCu versus the control (OceanBlu). The overall crude incidences of new IMI for 2 wk at risk were 4.9 and 7.3% for the ZkinCu and OceanBlu groups, respectively. The predominant organisms recovered from quarters with new IMI were Streptococcus uberis, Corynebacterium spp., and coagulase-negative staphylococci in both the ZkinCu and OceanBlu groups. The risk of infection in the OceanBlu group was higher (ß = 0.644; 95% confidence interval = 0.05-1.22). The interaction of treatment by week was not significant. The new IMI rate estimates (95% confidence interval) for ZkinCu and OceanBlu were 1.7% (0.8-2.5) and 3.2% (1.7-4.7), respectively. One novel aspect of this study is that it was one of the first commercial noninferiority trials to evaluate a new pre- and post-milking teat disinfectant in a dairy herd with an automatic milking system. The experimental teat disinfectant ZkinCu, evaluated in this field trial with naturally occurring IMI, showed noninferiority relative to the positive control for the prevention of new IMI. This study was conducted in a herd with an automatic milking system, and the results are applicable to herds with similar characteristics. Additional studies are needed to ensure reproducibility under different management conditions.

Identificación de especies de Mycoplasma y de Ureaplasma diversum en rodeos lecheros de Argentina / Identification of species of Mycoplasma and Ureaplasma diversum from Argentinian dairy herds

Varias especies de Mycoplasma y Ureaplasma diversum pueden causar enfermedades en el ganado bovino lechero, asociadas o no a manifestaciones clínicas. En nuestro país, ha sido detectada la presencia de solo tres especies de este grupo hasta el momento: Mycoplasma bovis, Mycoplasma californicum y Mycoplasma canadense. El objetivo del presente trabajo fue identificar otras especies de la familia Mycoplasmataceae. Se estudiaron treinta y cinco aislamientos compatibles con Mycoplasma spp. obtenidos a partir de diferentes muestras de bovinos, con o sin sintomatología clínica, provenientes de ocho rodeos ubicados en las provincias de Santa Fe, Córdoba, Buenos Aires y San Luis. Mediante el uso de reacciones en cadena de la polimerasa (PCR) específicas de especie se identificaron Mycoplasma bovigenitalum, Mycoplasma alkalescens, Mycoplasma bovirhinis y U. diversum, y mediante la amplificación y posterior secuenciación del espacio intergénico 16-23S ARNr se identificaron Mycoplasma arginini y M. californicum. La identificación de estas especies por primera vez en nuestro país es un hecho de Argentina relevancia, que representa un importante avance en el conocimiento para incluir estos patógenos en el diagnóstico diferencial de determinadas entidades clínico-patológicas de los bovinos de Argentina.

Several species of Mycoplasma and Ureaplasma diversum can cause diseases in dairy cattle, which can be associated or not with clinical manifestations. In our country, the presence of Mycoplasma bovis, Mycoplasma californicum and Mycoplasma canadense has been detected, being the only mycoplasma species identified so far. The objective of this study was to identify other species of the Mycoplasmataceae family. Thirty-five Mycoplasma spp.-like isolates obtained from different samples from cattle, with or without clinical symptoms, from eight herds located in the provinces of Santa Fe, Cordoba, Buenos Aires and San Luis were utilized in the present study. Through the use of species-specific polymerase chain reactions (PCR) Mycoplasma bovigenitalium, Mycoplasma alkalescens, Mycoplasma bovirhinis and U. diversum were identified and through amplification and further sequencing of the 16-23S rRNA intergenic spacer regions, Mycoplasma arginine and M. californicum were identified. The identification of these species represents an important advance in knowledge in order to include these pathogens in the differential diagnosis of certain clinical and pathological entities of cattle from Argentina.

[Identification of species of Mycoplasma and Ureaplasma diversum from Argentinian dairy herds]. / Identificación de especies de Mycoplasma y de Ureaplasma diversum en rodeos lecheros de Argentina.

Several species of Mycoplasma and Ureaplasma diversum can cause diseases in dairy cattle, which can be associated or not with clinical manifestations. In our country, the presence of Mycoplasma bovis, Mycoplasma californicum and Mycoplasma canadense has been detected, being the only mycoplasma species identified so far. The objective of this study was to identify other species of the Mycoplasmataceae family. Thirty-five Mycoplasma spp.-like isolates obtained from different samples from cattle, with or without clinical symptoms, from eight herds located in the provinces of Santa Fe, Cordoba, Buenos Aires and San Luis were utilized in the present study. Through the use of species-specific polymerase chain reactions (PCR) Mycoplasma bovigenitalium, Mycoplasma alkalescens, Mycoplasma bovirhinis and U. diversum were identified and through amplification and further sequencing of the 16-23S rRNA intergenic spacer regions, Mycoplasma arginine and M. californicum were identified. The identification of these species represents an important advance in knowledge in order to include these pathogens in the differential diagnosis of certain clinical and pathological entities of cattle from Argentina.

Quantification of antimicrobial usage in dairy cows and preweaned calves in Argentina.

Antimicrobials are commonly used to treat diseases in dairy cows and in calves. In Argentina information about veterinary antimicrobial usage at herd level is still limited. The purpose of this study was to quantify antimicrobial usage at herd level of most used drugs to treat lactating cows (LC) and preweaned calves (PWC). Prevalence of diseases and antimicrobial drug usage in 18 milking herds and in 11 calves rearing units (CRUs) were assessed through a survey. Drug usage (DU) at herd level was estimated through a standardized indicator, the number of Defined Daily Doses Animal (DDDA) per year, considering a standardized body weight of 600kg for LC and of 60kg for PWC. Pearson correlation coefficients were estimated and used to evaluate the association of LC herd size and milk yield (kg/day) with each disease prevalence and with DU. Student t-Test was used to compare disease prevalence and DU with various management practices in CRUs. Clinical mastitis was the most prevalent disease followed by foot, uterine and respiratory diseases in adult dairy cows. More involvement of veterinarians in treatment decisions was observed in larger dairy herds. Most used antimicrobials were beta-lactams and aminoglycosides. Especially for intramammary compounds, there was a trend towards multidrug formulations. The median DU was 5.21 DDDA/LC/year (range=2.88-10.88), the intramammary usage for clinical mastitis (IM-CM) and dry cow therapy (IM-DRY), representing 85.4% of total drug usage. No significant correlations were observed between herd size or milk yield with disease prevalence and with DU for all considered uses (IM-CM, IM-DRY and parental (PAR)). Enteritis was reported in all CRUs, followed by respiratory disease (91%), and omphalophlebitis (3%). The median DU for all drugs used at CRU level was 0.49 DDDA/PWC/year. Prevalences and DU for treatment of enteritis and respiratory diseases did not differ significantly between rearing systems, colostrum management or permanence time of the calf with the mother after birth. Although it was possible to estimate the drug usage at farm level, it was observed that the diversity of drug compounds and dose heterogeneity presented by the various manufacturers made the calculation of those indicators complex.

Comparison of phenotypic tests for detecting penicillin G resistance with presence of blaZ gene in Staphylococcus aureus isolated from bovine intramammary infections.

Few studies have described the relationship between genotypic and phenotypic methods for detecting penicillin resistance in Staphylococcus aureus isolated from bovine intramammary infection (IMI). Six phenotypic methods for penicillinase detection were compared with a genotypic method testing the presence of the ß-lactamase gene blaZ in Staph. aureus (n = 150) isolated from bovine IMI. Highest sensitivities and specificities were observed for disk diffusion (DD) (93 and 97.4%), minimum inhibitory concentration (MIC) (90.3 and 97.4%), Cefinase™ (85.9 and 97.4%) and Diatabs™ (85.7 and 98.7%). The estimated cut-off points estimated in the present study can be considered close to the ones indicated by CLSI (2013). The molecular detection of blaZ gene is the only method that may indicate the real or potential capacity of producing ß-lactamase in Staph. aureus. Considering that from a clinical standpoint a false negative result from a phenotypic test is the most unfavourable situation, a combination of standard DD with Diatabs™ or Cefinase™ should be performed by routine mastitis laboratories to minimise false negative results.

Detection of Mycoplasma canadense and Mycoplasma californicum in dairy cattle from Argentina.

Different species of Mycoplasma can affect bovine cattle, causing several diseases. PCR sequencing and further analysis of the 16S-23S rRNA ITS region have shown a significant interspecies variability among Mollicutes. Sixteen suspected isolates of Mycoplasma spp. obtained from milk samples from dairy herds were amplified (16S-23S rRNA ITS region). Fourteen out of those 16 suspected Mycoplasma spp. isolates were PCR-positive. To confirm the identity of Mycoplasma bovis, these 14 isolates were tested by another species-specific PCR. Seven of the isolates rendered a positive result. The products of 16S-23S rRNA ITS PCR from one isolate that was identified as M. bovis and from two other isolates, identified as non- M. bovis were randomly selected, sequenced and analyzed. The three sequences (A, B and C) showed 100% similarity with M. bovis, Mycoplasma canadense and Mycoplasma californicum respectively.

Detección de Mycoplasma canadense y Mycoplasma californicum en ganado lechero de Argentina / Detection of Mycoplasma canadense and Mycoplasma californicum in dairy cattle from Argentina

Diferentes especies del género Mycoplasma pueden afectar al ganado bovino y causar varias enfermedades. La técnica de PCR, secuenciación y posterior análisis de la región ITS 16S-23S ARNr ha mostrado que existe una importante variabilidad interespecies entre Mollicutes. Se realizó la amplificación (región ITS 16S-23S ARNr) de 16 aislamientos sospechosos de corresponder a alguna especie de Mycoplasma, que habían sido obtenidos de muestras de leche provenientes de rodeos lecheros. Catorce de esos aislamientos fueron PCR positivos. Para confirmar la identidad de Mycoplasma bovis, dichos aislamientos fueron evaluados por otra PCR especie-específica. Siete aislamientos dieron un resultado positivo. Los productos de la PCR de la ITS 16S-23S ARNr de un aislamiento identificado como M. bovis y de otros dos aislamientos identificados como no-M. bovis fueron seleccionados al azar, secuenciados y analizados. Las tres secuencias (A, B y C) mostraron 100 % de similitud con cepas de M. bovis, Mycoplasma canadense y Mycoplasma californicum, respectivamente

Different species of Mycoplasma can affect bovine cattle, causing several diseases. PCR sequencing and further analysis of the 16S-23S rRNA ITS region have shown a significant interspecies variability among Mollicutes. Sixteen suspected isolates of Mycoplasma spp. obtained from milk samples from dairy herds were amplified (16S-23S rRNA ITS region). Fourteen out of those 16 suspected Mycoplasma spp. isolates were PCR-positive. To confirm the identity of Mycoplasma bovis, these 14 isolates were tested by another species-specific PCR. Seven of the isolates rendered a positive result. The products of 16S-23S rRNA ITS PCR from one isolate that was identified as M. bovis and from two other isolates, identified as non- M. bovis were randomly selected, sequenced and analyzed. The three sequences (A, B and C) showed 100% similarity with M. bovis, Mycoplasma canadense and Mycoplasma californicum respectively

Detection of Mycoplasma canadense and Mycoplasma californicum in dairy cattle from Argentina. / Detection of Mycoplasma canadense and Mycoplasma californicum in dairy cattle from Argentina.

Different species of Mycoplasma can affect bovine cattle, causing several diseases. PCR sequencing and further analysis of the 16S-23S rRNA ITS region have shown a significant interspecies variability among Mollicutes. Sixteen suspected isolates of Mycoplasma spp. obtained from milk samples from dairy herds were amplified (16S-23S rRNA ITS region). Fourteen out of those 16 suspected Mycoplasma spp. isolates were PCR-positive. To confirm the identity of Mycoplasma bovis, these 14 isolates were tested by another species-specific PCR. Seven of the isolates rendered a positive result. The products of 16S-23S rRNA ITS PCR from one isolate that was identified as M. bovis and from two other isolates, identified as non- M. bovis were randomly selected, sequenced and analyzed. The three sequences (A, B and C) showed 100

similarity with M. bovis, Mycoplasma canadense and Mycoplasma californicum respectively.