Bacteriological cure rate and changes in milk composition in mastitis vaccinated ewes affected with subclinical mastitis.

AIM: The aim of this study was to investigate the effects of using a commercially-available polyvalent mastitis vaccine on the bacteriological cure rate of existing subclinical mastitis in Awassi sheep. MATERIALS AND METHODS: A total of 164 lactating ewes were divided into two main groups according to udder health and milk somatic cell count (SCC): Group 1=normal (N; n=80) and Group 2=subclinical mastitis (SC; n=84). Each group was then subdivided randomly into two treatment groups: N vaccinated (Nvax; n=38), N non-vaccinated (Nnvax; n=42), SC vaccinated (SCvax; n=42), and SC non-vaccinated (SCnvax; n=42). The vaccine was administered as per manufacturer's recommendations. Milk samples were collected aseptically from all ewes before vaccine administration (T0) and again on days 28 (T2) and 42 (T3) of the experiment. RESULTS: In the SC group, the bacteriological cure rates in vaccinated and non-vaccinated ewes were 76% and 69%, respectively. In N group, the new intramammary infection rates in vaccinated and non-vaccinated ewes were 48% and 50%, respectively. Vaccination of normal ewes resulted in a significant (p<0.05) reduction in bacterial growth rate both at day 28 and day 42 of the study. The prevalence of new intramammary infection rate in Nvax ewes on days 28 and 42 was 19% and 20%, respectively. The prevalence of new intramammary infection rate in Nnvax group on days 28 and 42 was 33% and 30%, respectively. In SCvax group, the bacterial growth rate on days 28 and 42 was 44% and 35%, respectively. In SCnvax group, the bacterial growth rate on days 28 and 42 was 27% and 32%, respectively. There was no statistically significant effect of vaccination on any of the studied milk composition parameters. CONCLUSIONS: This is a preliminary study that indicated a possible protective effect of vaccination against mastitis in sheep. Further, case-controlled studies are indicated to estimate the level of immunity this vaccine provides to vaccinated sheep.

Effect of androgen ablation on prostatic cell differentiation in dogs.

This study was conducted to gain further insight into the role of androgen in maintaining a balanced prostate gland growth in dogs. Effects of castration on prostatic cell were assessed by comparing the expression level of high molecular weight cytokeratin (HMW), alpha-actin, and vimentin in intact and castrated dogs. Mature dogs were castrated while they were under general anesthesia and were killed after 1 month. Mature prostate gland structures from intact dogs are characterized by the presence of differentiated columnar secretary epithelial cells and progenitor basal cells that are located within acini and ducts embedded in a thin fibromuscular tissue. Basal cells were distinguished from secretory epithelial cells by HMW cytokeratin immunostaining, which is expressed specifically by basal cells but not by epithelial cells. Castration-induced secretory epithelial cell death, leave the basal cells intact to form a continuous layer lining the atrophied acini. However, the survived basal cells lost their capacity to differentiate to secretory epithelial cells. In addition, androgen ablation induced remarkable reorganization of the cellular components of the fibromuscular compartment. In intact dogs, this compartment of prostate gland is composed mainly of differentiated smooth muscles and scattered mesenchymal muscles as reflected by the high and low actin and vimentin expressions, respectively. Castration for 1 month induced a progressive shift toward mesenchymal cells, which appeared to occupy most of the fibromuscular compartment. Based on these findings, it appears that androgen acts to maintain a steady state of prostate gland by driving the differentiation of prostatic cells and by maintaining its fully differentiated state.

Hoof wall wound repair.

REASONS FOR PERFORMING STUDY: Surgical stripping of the hoof wall results in a wound that heals remarkabley well. In contrast, lamellae recovering from laminitis are often deformed. Investigating lamellar wound healing may aid understanding of laminitis. OBJECTIVES: To document temporal changes in the lamellar basement membrane (BM), dermis and epidermis after surgery. METHODS: Wall strips were made in the dorsal hoof wall midline of 6 mature horses. Immunohistochemistry was used to document changes in the basement membrane (BM) and detect proliferation of epidermal cells in lamellar tissues harvested at intervals. A conforming metal plate was screwed to the hoof wall to maintain alignment of the wound edges. RESULTS: Wall stripping caused lamellar tips to snap and remain behind in the dermis along with the majority of the lamellar BM and some lamellar basal cells. Three days later the BM was intact and new lamellae had been reconstructed by proliferation of surviving epidermal cells. By 5 days the surface of the stripped zone was covered with yellow epidermis that subsequently thickened and hardened. Eventually the hoof wall deficit was replaced by new wall growing down from the coronet. The conforming metal plate and post operative analgesic ensured minimal lameness. CONCLUSIONS AND POTENTIAL RELEVANCE: In wall stripped lamellae the BM survives virtually intact and is used as a template for proliferating cells, from snapped-off lamellar tips, to migrate and quickly achieve repair to near normality. In laminitis epidermal dysadhesion and lamellar BM destruction occurs and lack of a functional BM template may explain the prolonged and abnormal repair of affected lamellae.

Epidermal cell proliferation in the equine hoof wall.

REASONS FOR PERFORMING STUDY: Current theories explaining how the hoof wall 'grows' and moves past the stationary distal phalanx are speculative and based on incomplete evidence. Movement in the lamellar region could occur by cell proliferation or an enzyme-based remodelling process. Since laminitis pathogenesis appears to involve increased transcription and activation of enzymes normally involved in tissue remodelling, it is important to know precisely which process dominates the lamellar region of the hoof.. OBJECTIVES: To investigate epidermal cell proliferation in the equine hoof wall and calculate a proliferative index (PI) for the coronet, lamellae and toe. METHODS: An analogue of thymidine, 5-bromo-2'-deoxyuridine (BRdU), was infused i.v. into 5 ponies. After tissue harvesting, BRdU (and therefore basal cell proliferation) was detected immunohistochemically using mouse anti-BRdU. PIs were calculated for the coronet and 10 levels of the dorsal hoof wall lamellae. RESULTS: The highest PIs (mean +/- s.e.) were in the coronet; 12.04% +/- 1.59 and proximal lamellae (7.13% +/- 1.92) and are therefore growth zones of the proximal hoof wall. PIs of more distal lamellae were 0.11% +/- 0.04 to 0.97% +/- 0.29; significantly lower (P = 0.05) than the lamellar growth zone. CONCLUSIONS: A 20-fold PI decrease between proximal and more distal lamellae suggests that the majority of the normal lamellae are nonproliferative and their main function is to suspend the distal phalanx within the hoof capsule. Remodelling within the hoof wall epidermal lamellae, which must occur as the hoof wall moves past the stationary distal phalanx, is a process not requiring epidermal cell proliferation. POTENTIAL RELEVANCE: A hoof lamellar epidermis that remodels using the same MMPs involved in laminitis pathogenesis implies that laminitis is a normal process out of control. Understanding MMP control and how the normal lamellar epidermis achieves this will help in the development of better laminitis preventative and treatment strategies.

Equine laminitis basement membrane pathology: loss of type IV collagen, type VII collagen and laminin immunostaining.

Disintegration of the basement membrane (BM) of the equine hoof lamellae and failure of the BM to remain attached to the basal cells of the secondary epidermal lamellae (SEL) is one of the earliest pathological events to occur in acute laminitis. Changes in the lamellar basement membrane were investigated by immunolabelling the key structural components of the BM, type IV collagen, type VII collagen and laminin in the lamellar BM of horses 48 h after the induction of laminitis. Lamellar tissues were harvested from 2 normal horses and 2 horses with acute laminitis. Immunostaining with antibody raised against human epitopes for type IV collagen, type VII collagen and laminin successfully stained the basement membranes of horse hoof lamellar tissues. Vascular tissue did not immunostain with type VII collagen antibody. Normal BM stained as a fine dark brown line and the lamellar BM was adhered to the basal cells of the SELs with no evidence of lamellar separation. At least 2 changes to the lamellar BM occurred in acute laminitis: loss of attachment of lamellar epidermal basal cells to their underlying BM and disintegration of the lamellar BM. In some sections from feet affected by acute laminitis, there was widespread separation of the SELs from their BM without loss of BM immunostaining and in others there was extensive loss of BM immunostaining. In lesions characterised by lamellar separation, the epidermal basal cells at the tips of the primary epidermal lamellae appeared to have slipped away from their BM and were an amorphous clump of epidermal cells devoid of immunostained BM. The BM from which they had separated remained in its original position in the dermis and was clearly outlined by all 3 antibodies. In other areas, however, virtually all the BM immunoreactivity at the PEL tips was absent. Only the occasional distorted SEL tip and fragments of BM retained sufficient immunostaining to allow anatomical identification. Numerous polymorphonuclear leucocytes (PMNs) invariably surrounded the tips of lamellae showing large scale loss of immunoreactivity and many PMNs had penetrated the lamellar BM and were within the epidermal compartments. PMNs were less frequent in the midlamellar region. Immunostaining of the BM of many SELs was absent in the midlamellar region. In some lamellae loss of BM immunostaining had occurred only at the bases of the SELs and fragments of immunostained BM were present in the zones of lysed BM suggesting that BM lysis was incomplete at the time of tissue fixation. In other lamellae, lysis of the BM was complete; there was no immunostained BM between SELs and the bulk of the epidermal cells of each PEL were an amorphous column of cells on either side of the central keratinised axis of the PEL. The lamellar BM which remained appeared as immunostained strands of unattached BM along the edges of the PDLs. Activation of BM degrading metalloproteinases (MMPs) occurs in acute laminitis and it seems likely that uncontrolled MMP activity is responsible for the loss and disorganisation of lamellar BM demonstrated in this study.