V. Bronzo

Machine-induced teat tissue reactions and infection risk in a dairy herd free from contagious mastitis pathogens

Machine-induced changes in teat thickness were measured in a randomly selected group of 22 cows from a commercial dairy herd consisting of 110 cows during an experiment lasting 1 year. Half the cows used were free from mastitis whereas the remainder had at least one quarter infected by environmental pathogens. Teats were classified according to a threshold change in teat end thickness of 5%. The relationships between quarter infection or teat duct colonization and teat end thickness changes induced by machine milking were investigated. Teats with greater than 5% change in thickness had significantly increased teat duct colonization and a slight, non-significant, increase in quarter infection. We conclude that machine-induced changes in teat end thickness are predisposing factors for teat duct colonization by environmental pathogens. This may provide an increased new infection risk, especially when hygiene is poor.

Ultrasound-guided cytology of spleen and liver: a prognostic tool in canine cutaneous mast cell tumor.

BACKGROUND In the clinical staging of cutaneous mast cell tumors (cMCT), the diagnosis of metastasis is controversial based on cytological examination of lymph nodes, spleen, liver, bone marrow, and blood. OBJECTIVES To define the prognostic role of ultrasound-guided cytology of spleen and liver in cMCT. The results of cytological evaluation were compared in relation with survival time. ANIMALS Fifty-two client-owned dogs with a diagnosis of cMCT. METHODS Selection of cases was based on cytological evaluation of liver and spleen to detect infiltration at distant sites. The Kaplan Meier method was used to compare survival in dogs with and without infiltration of spleen and liver (log-rank test P < .05). RESULTS Ten dogs with cMCT had mast cell infiltration of spleen, liver, or both and 4 of these dogs had involvement of the regional lymph nodes. The majority of dogs had 2 or more ultrasonographically abnormal findings simultaneously in spleen and liver. Nine dogs had grade II cMCT, and 1 had grade III cMCT. Dogs with positive evidence of mast cell infiltration to spleen, liver, or both had shorter survival times (34 versus 733 days) compared with dogs negative for mast cell infiltration at distant sites. CONCLUSION AND CLINICAL IMPORTANCE Dogs with evidence of mast cell infiltration at distant sites have a shorter survival times than dogs without evidence of infiltration at distant sites. This study suggests that cytology of spleen and liver is indicated either for ultrasonographically normal or for ultrasonographically abnormal spleen and liver in dogs with cMCT.

Study on the relationship between milk immune factors and Staphylococcus aureus intramammary infections in dairy cows.

The distribution of Staphylococcus aureus within herds seems to be related to interactions among the shedding characteristics of the bacteria, their pathogenicity and mammary gland immune status. The aim of the present study was to investigate the relationships between selected mammary gland immune factors and intramammary infections associated with Staph. aureus. Overall, 70 cows from five commercial dairy herds were included in the study and quarter milk samples were assessed using bacteriological and cytological tests. We evaluated differential cell count, lysozyme concentration, N-acetyl-beta-glucosaminidase (NAGase) activity, cell viability and respiratory burst activity in randomly chosen quarter milk samples from each cow. Staph. aureus intramammary infection elicited different responses in the mammary gland immune defences investigated. Polymorphonuclear leucocytes (PMN) as a proportion of total somatic cells in milk, cell viability and NAGase activity were higher in infected quarters, while the proportions of macrophages and lymphocytes, respiratory burst activity and lysozyme levels were lower. Mean values differed among herds, but the differences were not significant. These changes were associated with Staph. aureus infection. The reduced respiratory burst activity together with the increase in the proportion of PMN suggests that both the number and activity of PMN could influence the susceptibility of the mammary gland to pathogens. Indeed, the logistic model adopted suggests that impairment of milk immune factors could be concurrent with the development of an infection.

Effectiveness of electroacupuncture analgesia compared with opioid administration in a dog model: a pilot study

BACKGROUND Although opioid analgesics are the usual drugs to treat post-surgical pain, acupuncture has also been demonstrated to relieve various pain syndromes. The present pilot study aims to investigate the efficacy of electroacupuncture compared with a conventional opioid compound, butorphanol, for postoperative pain treatment in dogs undergoing elective ovariohysterectomy. METHODS Twelve dogs were randomly allocated into two groups. Dogs received either electroacupuncture stimulation (16 and 43 Hz) at Shen Shu, Chang Shu, He Gu, Tai Yuan, Zu San Li, Yang Ling Quan, and Bai Hui acupoints, while control dogs were treated with butorphanol. Cardiovascular and respiratory parameters were recorded for both groups during operation. Plasma β-endorphin concentrations were evaluated before surgery (baseline) and up to 24 h later. For each dog, pain was measured according to a dedicated subjective pain scoring system. RESULTS Plasma β-endorphin levels in dogs receiving electroacupuncture increased significantly against baseline values after 1 and 3 h after surgery. Moreover, the end-tidal isoflurane concentration needed for second ovary traction was significantly lower in acupuncture-treated dogs than control animals. All animals having electroacupuncture experienced prolonged analgesia, over 24 h at least, while four out of six dogs treated with butorphanol needed post-surgical ketorolac and tramadol supplementation to their pain relief. CONCLUSIONS The results obtained from the present investigation showed some evidence for electroacupuncture as an alternative technique to provide postoperative analgesia in dogs.

Efficacy of vaccination on Staphylococcus aureus and coagulase-negative staphylococci intramammary infection dynamics in 2 dairy herds.

The aim of this study was to evaluate vaccine efficacy of a commercial vaccine (Startvac, Hipra Spain) aimed at reducing intramammary infections (IMI) with Staphylococcus aureus and coagulase-negative staphylococci under field conditions. During the 21-mo duration of the study, 1,156 lactations from 809 cows were enrolled in 2 herds. During the first phase of the trial, all cows that were due to calve were vaccinated until approximately 50% of cows in the milking herd were vaccinated (at ~6mo). At that point, when 50% vaccination coverage was reached, cows that were due to calve were randomly assigned to be vaccinated or left as negative controls. Cure rate, rate of new infection, prevalence, and duration of infections were analyzed. Vaccination resulted in a moderate reduction in incidence of new staphylococcal IMI and a more pronounced reduction in duration of IMI associated with reduction of the basic reproduction ratio of Staph. aureus by approximately 45% and of coagulase-negative staphylococci by approximately 35%. The utilization of vaccine in combination with other infection-control procedures, such as excellent milking procedures, treatment, segregation, and culling of known infected cattle, will result in an important reduction in incidence and duration of intramammary staphylococcal infections.

Double pelvic osteotomy for the treatment of hip dysplasia in young dogs

UNLABELLED The aim of this study was to evaluate the feasibility of the double pelvic osteotomy (DPO) (osteotomy of the ilium and pubis) to treat clinical cases of hip dyplasia in young dogs instead of performing a triple pelvic osteotomy (TPO) (osteotomy of the ilium, pubis, and ischium). Candidates for DPO were 4.5- to nine-month-old dogs with coxofemoral joint subluxation and laxity, indicative of susceptibility to future development of severe hip dysplasia. The angle of reduction (AR) and angle of subluxation (AS) with Ortolanis sign, Norberg angle (NA), percentage of femoral head (PC) covered by the acetabulum, and the pelvic diameters and their relationships were measured clinically and radiographically before and after surgery. The surgical technique was similar to the TPO technique, but excluded ischiatic osteotomy. A DPO was carried out in 53 joints of 34 dogs; AR and AS values immediately postoperatively and at the one- and two-month follow-up examinations were significantly lower than the preoperative values (p <0.01). The complications encountered were mainly represented by implant failure (3.5%), partial plate pull-out (9.4%), and incomplete fracture of the ischial table (7.5%). Changes in PC and NA values obtained immediately after surgery and at the first and second follow-up examinations were significantly greater (p <0.01 both) than values obtained before surgery. Sufficient acetabular ventroversion was achieved to counteract joint subluxation and the modifications of AR and AS. The NA and PC direct postoperative values reflected a significant improvement in the dorsal acetabular coverage. CLINICAL RELEVANCE Restoration of normal joint congruity (PC from 50 to 72%) and maintenance of the pelvic geometry without pelvic narrowing were the most intriguing features of DPO. The complications observed were greatly reduced when using dedicated DPO plates. Based on our experience, the morbidity after unilateral and bilateral DPO was lower than after TPO because elimination of the ischiatic osteotomy allowed for increased stability of the pelvis. The surgical technique of DPO was a little more demanding than TPO because of the difficulty in handling and rotating the acetabular iliac segment, but this difficulty was offset by elimination of ischial osteotomy.

Alpha1-acid glycoprotein is contained in bovine neutrophil granules and released after activation

The present study was designed to investigate the capability of bovine neutrophil granulocytes to produce the minor acute phase protein alpha(1)-acid glycoprotein (AGP, Orososmucoid). Bovine neutrophils contain a high MW (50-60kDa) AGP isoform (PMN-AGP), as determined by Western blotting and confirmed by fluorescence microscopy. The presence of AGP in bovine neutrophils has been confirmed by fluorescence immunocytometry. In addition, bovine neutrophils contain also a 42-45kDa isoform, which has the same MW as plasma-, liver-delivered, AGP. cDNA sequence of plasma- and PMN-AGP revealed that (i) the two proteins are products of the same gene; (ii) the differences in molecular weight are due do different post-translational modifications. This result was confirmed by deglycosylation of the two glycoforms. Exocytosis studies showed that isolated neutrophils exposed to several challengers, including Zymosan activated serum (ZAS) and phorbol 12-myristate 13-acetate (PMA), which mimic the inflammatory activation, released PMN-AGP as early as 15min. AGPs mRNA is physiologically expressed by mature resting neutrophils. Real-time PCR on LPS, ZAS and PMA challenged cells revealed that the level of expression apparently does not increase after inflammatory activation. Collectively, the findings reported in this paper proved that PMN-AGP: (i) is a hyperglycosylated glycoform of plasma AGP, (ii) is stored in granules, and (iii) is released by neutrophils in response to activation. Due to its anti-inflammatory activity, PMN-AGP may work as a fine tuning of the neutrophils functions in the inflammatory focus, i.e. it can reduce the damages caused by an excess of inflammatory response.

Field study on the relationship between teat thickness changes and intramammary infections

The aim of this study was to describe the results of teat thickness measurement applied routinely in three commercial dairy herds and to evaluate the influence of machine-induced teat thickness changes on intramammary infection and the frequency of new infection. A total of 1018 fore milk samples and the same number of teat apex measurements have been evaluated. Overall, relative teat thickness changes were normally distributed (mean -0.16%, SD 10.15%), while a specific pattern could be observed within herds. Increases in teat thickness of > 5% were significantly associated with infection and new infection (odds ratio > 1), but the association was not significant when teat thickness decreased by more than 5%. When results were classified according to aetiology, analysis showed that coagulase-negative staphylococcal infections were significantly associated with both increases and decreases in teat thickness numerically greater than 5%.

Escherichia coli lipopolysaccharides and Staphylococcus aureus enterotoxin B differentially modulate inflammatory microRNAs in bovine monocytes.

MicroRNAs (miRNAs) are a family of regulatory molecules involved in many physiological processes, including activation of cells of the immune system. This study investigated the effect of Escherichia coli lipopolysaccharide (LPS) and Staphylococcus aureus enterotoxin B (SEB) on the expression of five miRNAs involved in the inflammatory response, including miR-9, miR-125 b, miR-155, miR-146 a and miR-223, in bovine CD14(+) cells (monocytes). Incubation of monocytes with SEB induced down-regulation of miR-155, miR-223 and miR-125 b, but not the anti-inflammatory miRNA miR-146 a. Conversely, incubation with LPS upregulated both miR-155 and miR-146 a. In vitro incubation of isolated CD14(+) bovine monocytes with LPS and SEB elicited different and opposite expression of miRNAs reportedly involved in inflammatory reactions.

Differential expression and secretion of alpha1-acid glycoprotein in bovine milk.

alpha1-Acid glycoprotein (AGP) is a lipocalin that is produced mainly by the liver and secreted into plasma in response to infections and injuries. In this study, we evaluated AGP isoforms that can be detected in bovine milk. We found that milk-AGP content is made up of at least two isoform groups, a low MW group (44 kDa) that is produced in the mammary gland (MG-AGP), and a higher MW group (55-70 kDa), that is produced by somatic cells (SC-AGP). Identical SC-AGP isoforms can be found both in milk and blood PMN cells. Analysis of the mammary tissue cDNA showed that the sequence of the MG-AGP isoform is identical to that of plasma AGP. Each group contains several proteins with different MWs and different isoelectric points, as shown by 2D-electrophoresis. The glycosylation patterns of these isoforms were analysed by means of specific lectin binding, to evaluate the degree of sialylation, fucosylation and branching. The MG-AGP glycan pattern was identical to plasma AGP produced by the liver. Several differences were detected, however, between plasma and SC-AGP isoforms, the most evident being the strong degree of fucosylation and the elevated number of di-antennary glycans in SC-AGP. Immunohistochemistry showed that AGP is found in all tissues that make up the mammary gland, but that it is most likely produced for the main part by the alveoli.