Scanning electron microscopy-aided observations on and therapy of teat canal infections

An examination of teat canal swabs established that 51 teat canals out of 68 quarters of machine-milked cows were colonized by Staphylococcus aureus. Only 31 of these quarters yielded milk from which S. aureus could be cultured, and 6 out of the 31 produced milk containing somatic cell counts in excess of 500 000/ml. No inhibitory substances could be detected in milk samples 12 h after 10 mg of sodium cloxacillin had been deposited in the test canal on 1-4 successive occasions. Teat canal swabs and milk sample cultures of the same quarters became and remained bacteriologically negative for at least a week after the last treatment. Six quarters, which according to the International Dairy Federation criteria were suffering from subclinical mastitis, became negative after local teat canal therapy. Scanning electron micrographs of one infected teat canal revealed the presence of cocci in depressions and crevices on the epithelial surface, suggesting that such cocci are not always flushed out into milk samples. Teat canal therapy should make a marked contribution to the control of bovine mastitis.     

Changes in the bovine teat canal during the nonlactating period and early lactation, as measured by teat canal impressions.

Changes in dimensions of impressions of the lumen of the teat canals of 13 cows were examined at 17 intervals during the nonlactating period and early lactation. Impressions were made of teats of 2 diagonally opposed quarters of each cow, using dental impression material. Impression length was measured and cross sections of the impressions at the proximal (distal to Furstenburg rosette), distal (proximal to the teat orifice), and middle (midway between the 2), portions of the teat canal were prepared. Cross sections were photographed and enlarged, and circumference and area were determined by use of planimetry. Effects of making repeated impressions during the nonlactating period and early lactation on new infection rates and somatic cell counts were also assessed. Mean length of teat canal impressions decreased between days 0 and 3 of involution and during the prepartum periods. Depending on the level from which they were taken, cross-sectional areas of impressions tended to increase or increased significantly during the period of involution and again in the prepartum period. Significant changes in cross-sectional area were not observed during early lactation. Changes in circumference of proximal, middle, and distal cross sections followed trends similar to area measurements, but were more variable and differences were less statistically significant. On the basis of our findings, we suggest that heightened susceptibility to new infection during mammary involution and the prepartum period may be attributable, in part, to changes in the patency of the teat canal. Making impressions repeatedly throughout the nonlactating period and early lactation did not affect the number of new intramammary infections.     

The prophylactic effect of a dry-cow antibiotic against Streptococcus uberis

The prophylactic use of a dry-cow antibiotic for reducing the incidence of mastitis due to Streptococcus uberis was studied in four seasonally calving dairy herds involving 378 cows. The treatment was a long-acting dry-cow antibiotic preparation administered immediately after the last milking of lactation. New intramammary infections were identified by comparing the bacteriological status of quarters at drying off with that after calving, or through manual udder palpation during the dry period. The administration of dry-cow antibiotic to uninfected quarters at drying off reduced the overall incidence of new infections with Streptococcus uberis from 12.3% for untreated quarters to 1.2% of quarters (p<0.01). The reduction was significant (p<0.01) for both dry-period and post-calving infections. The susceptibility of uninfected quarters to new infection by Streptococcus uberis appeared to be unrelated to the infection status of a cow at drying off. Clinical infections during the dry period were most prevalent (97%) in quarters identified as having open teat canals. Fewer open teat canals (p<0.05) were observed among antibiotic treated quarters over the first 4 weeks of the dry period. Treated quarters had a lower (p<0.05) incidence of new clinical infection during the ensuing lactation and lower somatic cell counts. This did not affect production levels of milk, milk fat or protein. The results clearly indicated a prophylactic benefit for the dry cow antibiotic treatment against new Streptococcus uberis infections during the dry period.     

The prophylactic effect of a dry-cow antibiotic against Streptococcus uberis

The prophylactic use of a dry-cow antibiotic for reducing the incidence of mastitis due to Streptococcus uberis was studied in four seasonally calving dairy herds involving 378 cows. The treatment was a long-acting dry-cow antibiotic preparation administered immediately after the last milking of lactation. New intramammary infections were identified by comparing the bacteriological status of quarters at drying off with that after calving, or through manual udder palpation during the dry period. The administration of dry-cow antibiotic to uninfected quarters at drying off reduced the overall incidence of new infections with Streptococcus uberis from 12.3% for untreated quarters to 1.2% of quarters (p<0.01). The reduction was significant (p<0.01) for both dry-period and post-calving infections. The susceptibility of uninfected quarters to new infection by Streptococcus uberis appeared to be unrelated to the infection status of a cow at drying off. Clinical infections during the dry period were most prevalent (97%) in quarters identified as having open teat canals. Fewer open teat canals (p<0.05) were observed among antibiotic treated quarters over the first 4 weeks of the dry period. Treated quarters had a lower (p<0.05) incidence of new clinical infection during the ensuing lactation and lower somatic cell counts. This did not affect production levels of milk, milk fat or protein. The results clearly indicated a prophylactic benefit for the dry cow antibiotic treatment against new Streptococcus uberis infections during the dry period.     

Teat canal closure in non-lactating heifers and its association with udder health in the consecutive lactation

Prevention of heifer mastitis is a key element in preventing mastitis. Since pathogenesis of heifer mastitis is not fully understood yet, combating it is arduous. The goal of this study was to examine whether the premature loss of the keratin plug of the teat canal represents an important risk factor for the development of heifer mastitis. 84 dairy heifers (German Holstein) from six high-yielding dairy herds in Northern Germany were examined between March 2005 and June 2006. Each quarter was examined clinically at least three times before calving. If the teat canal of a quarter was open, secretion samples were collected for bacteriological analysis. After calving, four quarter samples were collected during the first lactation. A rapid increase of the proportion of open teat canals was detected in the preparturition period. No teat canals were open till 80 days before parturition, while 60% of the teat canals were open at 60 days before parturition. A time-scheduled infection pattern was observed: during the days 90-61, 60-31, and 30-0 before calving, skin inhabitants (coagulase-negative staphylococci, coryneform bacteria), cow-related, and environment-related microorganisms prevailed, respectively. A total of 77% of intramammary infections encountered at parturition had established previously. No significant association between the duration of open teat canals before calving and the udder health post partum was found. However, the incidence of clinical mastitis during first lactation was influenced largely by the duration of infection ante partum and the mastitis pathogen involved. This study shows that a high proportion of teat canals already opens several months before calving and that opening of the teat canal before calving is an important factor in the aetiology of heifer mastitis.     

Resistance mechanisms of the bovine udder: new implications for mastitis control at the teat end

Investigations on bovine teat-end defenses and their role in mastitis control were reviewed. Alteration of teat canal keratin by method of intramammary drug infusion through the teat canal influenced the number of new infections. At the beginning of the nonlactating period, 2 methods of administering antibiotic were studied: full insertion of treatment syringe cannulas into teat cisterns and expelling contents and partial insertion of cannulas into the distal 2 to 3 mm of teat canals and slowly infusing contents. Partial cannula insertion reduced new infections by 50% and is an easily adopted management tool to reduce prevalence of mastitis. Intramammary devices used during lactation and the nonlactating period increased leukocyte concentrations in mammary secretions. An abraded polyethylene coil device reduced clinical mastitis and increased milk yield. Staphylococci colonize teat canal keratin and lacteal secretions of dairy heifers as early as 9 months of age, leading to intramammary infection at time of calving and persisting into lactation. Subsequent somatic cell counts are associated with milk production losses. Previously, such infections were shown to be associated only with older, mature animals.     

Bovine serum albumin and cell counts in the diagnosis of subclinical udder infection

Summary

Puncture of the milk cisterns was performed in 120 bacteriologically positive quarters of forty‐seven lactating dairy cows on three farms. This method was used to determine whether the existing infection was an infection of the teat canal or one of the udder. The results were related to the concentration of bovine serum albumin (BSA) and the cell count in the milk. Of the bacteriologically negative quarters, both BSA levels (in 91 per cent of the quarters the BSA concentration was 0.20 mg. per ml. of milk or less) and cell counts (92 per cent contained less than 500,000 cells per ml. of milk) were low. In cases of udder infection with primary pathogenic bacteria there was a marked increase in cell count (90 per cent more than 500,000 cells per ml. of milk), whereas the increase in BSA was rather small (51 per cent still contained 0.20 mg. BSA per ml. of milk or less). While the difference in cell counts of milk from quarters with udder infections and teat canal infections with primary pathogenic bacteria was significant, the difference between the BSA levels of these two groups was not. Therefore, the cell count supplies more reliable information than does the BSA level of the milk. Of all infections, 23 per cent were found to be infections of the teat canal.     

Teat characteristics in relation to animal temperament during milking in buffaloes,and comparison of buffalo and cow teat morphology

This study aimed to examine the teat characteristics in relation to the animal temperament during milking in the Anatolian buffaloes using ultrasonographic, histomorphological and immunohistochemical methods. The teat canal length (TCL), teat wall thickness (TWT), teat cisternal diameter (TCD), teat diameter (TD), teat length (TL), and teat circumference (TC) values in docile (n = 5) and nervous (n = 7) buffaloes were measured at the 0th, 3rd and 6th minute of stimulation. In additional experiments, comparative histomorphology and immunohistochemical examinations of buffalo (n = 7) and cow teats (n = 8) were performed. It was determined that post-stimulation mean TCL values were significantly higher in nervous buffaloes than those of teats in docile buffaloes (p < .05). A significant positive correlation between TCD and TD, TL and TC in both docile and nervous buffaloes was noted (p < .05). Unlike nervous buffaloes where only 3/14 teat canals were open by 3rd minute of milking stimulation, almost all (9/10) teat canals were observed opened in docile buffaloes. There were fewer muscle but more collagen bundles in buffalo teats compared with cow teats. It seems that temperament of animal during milking effects the milking efficiency, and in nervous buffaloes, probably the stimulation alone may not be sufficient for opening of the teat canal and hence achieve complete milking.     

The prophylactic effect of a teat sealer on bovine mastitis during the dry period and the following lactation

AIM: To determine the prophylactic efficacy of a teat sealer, administered at drying off, in reducing new intramammary infections in the dry period and the following lactation. METHODS: A total of 528 cows with late lactation somatic cell counts <2 00,000 cells/ml was identified in three commercial herds. Of these, bacteriological examination showed 482 cows were uninfected in all four quarters and 46 were infected in only one quarter. At drying off, uninfected quarters were randomly allocated to the following treatments: no infusion (negative controls), infusion with a bismuth subnitrate based teat sealer, infusion with teat sealer + antibiotic, or infusion with a cephalonium-based dry cow antibiotic (positive control). New infections were identified during the dry period by periodic udder palpations and at calving by bacteriological culture. RESULTS: All three infused treatments reduced the incidence of new intramammary infections due to Streptococcus uberis, both during the dry period and at calving, by about 90% (p <0.01). The majority of the infections were due to Streptococcus uberis. For all treatments, a 50% lower incidence of clinical mastitis over the first 5 months of the ensuing lactation was reported by farmers. X-ray imaging of 19 teats showed that the teat sealer material was retained, at least in part, in the lower teat sinus over about 100 days of the dry period. CONCLUSIONS: Closure of the teat canal from day one of the dry period as achieved by the teat sealer was as effective in reducing new dry period infections as the infusion of a long-acting dry cow antibiotic formulation. The lower incidence of new infections in the ensuing lactation among the infused quarters implies that fewer subclinical infections persisted from the dry period. Use of teat sealers at drying off appears to offer the same prophylactic efficacy as the dry cow antibiotic approach.     

Chronic obstructive mastitis in the camel. A clinicopathological study

Unilateral chronic mastitis in three she camels was due to obstruction of the teat canal by keratin. This lead to dilatation of the ducts, retention of milk and secondary bacterial infection. The teat canals and dilated ducts were lined by stratified squamous epithelium. There was excessive periductal fibrosis. Pasteurella hemolytica was isolated from one animal and Staphylococcus aureus from another. The fluid from the third animal was sterile. The condition was treated successfully by surgical amputation of the affected halves of the udder.     

The susceptibility of bovine udder quarters colonized with Corynebacterium bovis to experimental infection with Staphylococcus aureus or Streptococcus agalactiae.

Twenty bovine udder quarters colonized with Corynebacterium bovis SR6 and 20 uncolonized quarters were challenged by inoculation of Staphylococcus aureus Newbould 305 (ATCC 29740) into the teat cistern. The percentage of infection in quarters colonized with C. bovis (50%) was significantly lower than that in controls (100%). By similar challenge no significant difference was observed between the percentage of infection with Streptococcus agalactiae ATCC 27956 in 33 quarters colonized with C. bovis (70%) compared to 33 controls (87.9%). A total of 37 quarters colonized with C. bovis and 37 control quarters were challenged with Staph. aureus Newbould 305 (ATCC 29740) and Maxi (ATCC 27543) and Strep. agalactiae (ATCC 27956) by exposure of the teat orifice. The percentage of teat ducts colonized with C. bovis which became infected with either pathogen was not different from that for controls.     

Influence of milking technique and lactation on the bovine teat by means of ultrasonographic examination

The teats of Brown Swiss and Austrian Simmental cows, divided into two groups, one milked by means of an automatic milking system, the other using a conventional milking parlour, were examined monthly by ultrasonography. Aim of the study was to compare the effects of two different milking machines upon the structures of the bovine teat canal and wall by ultrasonography and thereby evaluate ultrasonography as a research tool for visualisation of different influences on the bovine teat. Length and thickness of the teat canal and teat wall thickness were measured and analysed. During lactation, teat canal length and thickness increased in both groups, teat canal length decreased in conventional milked cows at the end of lactation. Shorter and narrower teat canals were observed in automatic milked cows. Differences between the groups in teat canal length and thickness were determined in early lactation. During lactation teat wall thickness showed a slight increase. Automatic milked cows displayed thinner teat walls than cows milked in the milking parlour. Teat morphology was influenced by the number and duration of lactations, milk yield, quarter of the udder and time and date of examination. It was concluded that the effect of the two different milking machines caused significant differences in bovine teat morphology and that ultrasonography proved to be an appropriate method for visualising influences of the milking technique on the bovine teat.     

Changes in the bovine udder quarters naturally infected by Corynebacterium bovis

Of 272 bovine udder quarters studied for mastitis, 19 of them naturally infected with Corynebacterium bovis alone, were compared with 16 others infected by C. bovis together with other bacteria and another 36 non-infected quarters. While there was no significant difference in milk somatic cell counts between the quarters infected by C. bovis alone and those affected by C. bovis together with other bacteria (33.37 +/- 20.28 X 10(3) and 33.86 +/- 23.18 X 10(3)/ml of milk, respectively), there was a significant difference between these and the non-infected quarters (5.60 +/- 3.23 X 10(3)/ml of milk). Microscopically, quarters infected by either C. bovis alone or C. bovis in combination with other bacteria had inflammatory changes in the teat cisterns, Furstenberg's rosettes and/or mammary parenchyma. The non-infected quarters had no changes. In all 82 quarters no pathological changes could be seen in the teat canals.     

Infrared thermography and ultrasonography to indirectly monitor the influence of liner type and overmilking on teat tissue recovery

Eight Danish Holstein cows were milked with a 1-mm thick specially designed soft liner on their right rear teat and a standard liner mounted under extra high tension on their left rear teat. Four of the animals were overmilked for 5 min. Rear teats were subjected to ultrasound examination on the first day and to infrared thermography on the second day. Teats were submersed in ethanol 20 min post-milking on the second day. Ultrasonography measurements showed that teat canal length increased by 30-41% during milking. Twenty minutes after milking, teats milked with modified standard liners still had elongated teat canals while teats milked with the soft liner were normalized. Overmilking tended to increase teat wall thickness. Approximately 80% of variability in teat canal length, from before teat preparation to after milking, could be explained by changes during teat preparation. Thermography indicated a general drop in teat temperature during teat preparation. Teat temperature increased during milking and continued to increase until the ethanol challenge induced a significant drop. Temperatures approached pre-challenge rather than pre-milking temperatures within 10 minutes after challenge. Teat temperatures were dependent on type of liner. Mid-teat temperatures post-challenge relative to pre-teat preparation were dependent on overmilking. Thermography and ultrasound were considered useful methods to indirectly and non invasively evaluate teat tissue integrity.     

Effect of damage to the teat end on the experimental induction of mastitis in dry cows with Corynebacterium pyogenes

The teat ends of 12 dry cows were contaminated with Corynebacterium pyogenes. To determine whether a pre-existing (an)aerobic bacterial infection of the udder was a predisposing factor for a C pyogenes mastitis they included infected and uninfected quarters. Anaerobic bacteria could not be found and mastitis was not induced. When the teats were contaminated with C pyogenes after the teat ends had been injured 30 of the quarters became infected, and anaerobic bacteria were demonstrated in many quarters.     

Role of leukotriene B4 in the pathogenesis of Klebsiella pneumoniae-induced bovine mastitis

Mastitis was induced in 4 lactating cows by inoculation of Klebsiella pneumoniae (10(7) organisms/ml) via the teat canal. Sterile isotonic saline solution (1 ml) was instilled into designated control quarters via the teat canal. Changes in milk leukotriene B4 and C4 (LTB4, LTC4) concentrations, milk somatic cell counts, and milk bovine serum albumin concentration were monitored over a 24-hour postinoculation period. Milk LTB4 concentration before inoculation in control quarters and quarters later to be infected was 376 +/- 45 and 326 +/- 56 pg/ml of milk, respectively. A significant (P less than 0.05) increase in milk LTB4 concentration in the infected quarters was first observed at postinoculation hour 6, and milk LTB4 concentration in infected quarters generally remained significantly high through postinoculation hour 14. Thereafter, milk LTB4 concentration in infected quarters was not significantly different from the concentration in control quarters. Measurable amounts of LTC4 were not detected in the milk of either control or infected quarters. Milk bovine serum albumin concentration in the infected quarters generally was high throughout the study, as were milk somatic cell counts. The results of this study suggested that LTB4 contributes to the pathogenesis of bovine mastitis.     

Cytologic observations of the bovine teat end

Cells infiltrating from the vasculature and histologic components of internal tissues of teats (mammary papilla) from noninfected udder quarters were studied, using light and electron microscopy. Morphometric analysis demonstrated a progressive increase in number of infiltrating cells from the distal teat cistern (sinus papillaris) to the junction of the Furstenberg's rosette (distal termination and convergence of mucosal folds lining the teat cistern) and the streak canal (ductus papillaris). Plasma cells contributed to cellular increases in subepithelial connective tissue and were the most prevalent infiltrating cell type. Plasma cells also penetrated the basal epithelial lining of the rosette area and occasionally migrated to the luminal surface near the squamocolumnar junction. Neutrophils and monocytes contributed to the increase in cells infiltrating the epithelial lining. Few infiltrating cells were observed in epithelium and underlying stroma of the streak canal. Cytologic comparison demonstrated a reduction in all cell types from lactating to involuting phases of lactation. Greater numbers of plasma cells, lymphocytes, and monocytes were observed in teat end tissues from quarters previously infected with Staphylococcus aureus.     

Teat lesions and their relationship to intramammary infections on small-scale dairy farms in Kiambu district in Kenya

Mammary gland quarters of 139 lactating dairy cows from small-scale dairy herds were examined visually and by palpation for teat lesions and by California mastitis test (CMT) and bacterial culture for subclinical mastitis. Teat lesions were observed in 97 teats. These included teat chaps (39.2%), teat papillomas (23.7%), teat erosions (22.7%), teat fistulae (5.1%), inverted teats (5.1%) and blocked teats (4.2%). According to the CMT, the prevalence of subclinical mastitis was 33.4% in all the mammary gland quarters, 71.0% in quarters with teat lesions and 24.5% in quarters without teat lesions. There was a significant (P < 0.01) association between teat lesions and the prevalence of subclinical mastitis. The mammary gland quarters with teat lesions were 7.2 times more likely to have a positive CMT (P < 0.01) and 5.6 times more likely to have bacterial organisms (P < 0.01) isolated from them than those without any teat lesions. The bacterial organisms most frequently isolated from the CMT-positive milk samples from both the mammary gland quarters with teat lesions and those without teat lesions were Staphylococcus aureus (50.0%), Streptococcus spp. (34.8%) and Arcanobacterium pyogenes (6.2%).     

Changes in the microscopic anatomy of the bovine teat canal during mammary involution

Changes in the microscopic anatomy of the bovine teat canal were examined during mammary involution. Morphometric analyses revealed a significant (P less than 0.05), temporary dilatation of the teat canal lumen on day 7 of the nonlactating period. Additionally, the teat canal epithelium physiologically atrophied as evidenced by decreased cross-sectional area and thickness during the first 30 days of the nonlactating period, significantly so (P less than 0.05) between days 0 and 7. This physiologic atrophy was due mainly to a reduction in area and thickness of the stratum granulosum and may have resulted from continuing keratinization, a process that led to increased thickness of the keratin layer and formation of a functional plug during later stages of involution. Changes in cells of the stratum granulosum indicated a decrease in the rate of epithelial cell maturation during involution. The mitotic index (percentage of basal cells in mitosis) of the teat canal epithelium decreased significantly (P less than 0.05) between days 0 and 7 of the nonlactating period. Bacteria, observed in histologic sections, appeared to colonize only certain regions of the keratin layer. Seemingly, changes in the teat canal during mammary involution may be important factors in changing susceptibility to new intramammary infection during the early and mid-nonlactating periods.     

Microscopic changes in infected bovine teats

Two hundred and fifty-two teats from the udder quarters of 68 dairy cows were studied by bacteriological examination of quarter milk samples and microscopic examination of the teats. Of 252 quarters, 169 teats showed inflammatory changes, while 83 were of normal appearance. The inflammatory changes consisted of either neutrophil infiltrations, mononuclear cell infiltrations and fibrosis, or a combination of these changes. The reaction was restricted to the teat cisterns and/or Fürstenberg's rosettes, but very rarely also in the teat canal. In 136 of 169 cases, the inflammatory reaction was associated with bacteria in the milk, while in 33 cases no bacteria could be recovered from milk, in spite of inflammatory changes in the teat structures. Only 36 of 83 of normal teats were bacteriologically positive.