Transmission of Mannheimia haemolytica from the tonsils of lambs to the teat of ewes during sucking

Objective of the work was to study whether Mannheimia haemolytica may be transmitted from the mouth of the lambs into the teat of the dam during sucking. We compared bacterial populations within the teat duct and milk of ewes immediately before and immediately after sucking by the lambs. Tonsils of lambs of the ewes were swabbed. M. haemolytica strain DAG21T recovered from a teat duct of a ewe was compared to strain DAG21R recovered from the tonsils of her lamb by using 16s rRNA sequencing. We used those two isolates and another one of known pathogenicity, for challenging ewes: (i) 2-mm deep into healthy teats, (ii) 2-mm deep into teats with chapping lesions or (iii) into the cistern of healthy mammary glands. Of samples collected before suckling, 20/792 were bacteriologically positive, and of those after, 50/792 were bacteriologically positive (P < 0.001); in 37 cases, a negative sample became positive. One M. haemolytica (DAG21T) was recovered after suckling from a teat duct of a ewe. The organism was isolated from 57/90 tonsillar swabs from lambs. Risk of infection of ewe’ teats was 0.004 throughout lactation, being greatest (0.021) during the 3rd week of lactation. The 16s rRNA sequences of strains DAG21T and DAG21R were identical over 1450 nucleotides. Phylogenetic analysis showed that the two isolates clustered together with isolates of M. haemolytica. Organism deposition into healthy teats caused subclinical mastitis; deposition into teats with lesions or directly into mammary gland caused clinical mastitis. When results of inoculation of the three strains were compared between them, statistical significance was always P > 0.9. Results provide clear evidence that suckling by lambs can lead to transmission of M. haemolytica into the teats of the ewes; the bacteria have the potential to cause mastitis if circumstances are favourable.     

Effects of machine-milking on the bacterial flora of teat duct and mammary gland of ewes

In total, 308 paired-samples of teat duct material and milk, were collected before and 50-70 min after machine-milking, from 30 ewes. Samples were processed bacteriologically. For analysis of results, we compared changes in bacterial isolation following milking, for duct and milk samples; statistical significance was assessed by the Sign Test. Bacteria were isolated from 18 (6%) duct and 19 (6%) milk samples collected before the milking procedure; respective figures after it, were 81 (26%) and 33 (11%). In 77 (25%) cases, bacteriological findings in the two duct samples of each pair were different; in seven cases bacteria were isolated only before, whilst in 70 cases bacteria were isolated only after milking (P < 0.005); respective results for milk samples were 26 (8%): 6 and 20 cases (P = 0.693). The majority of bacterial isolates were staphylococci, accounting for 63% of 99 isolates. The milking procedure predisposes to entrance of bacteria into the teat duct; however, increased bacterial isolation from the teat did not result to increased mammary infections, likely as a consequence of defence mechanisms present in healthy teats.     

Effects of lamb sucking on the bacterial flora of teat duct and mammary gland of ewes.

We aimed (i) to determine differences in bacterial flora of teat duct and mammary gland of ewes before and after suckling, (ii) to evaluate factors potentially affecting those. We collected samples of teat duct material and mammary secretion from 11 ewes immediately before and after sucking by lambs, as well as 120 min later. We processed samples bacteriologically and compared changes in infection by the Sign Test. We isolated bacteria from 3.5% duct and 1.5% secretion samples before suckling. Respective figures post-suckling were 10.6% and 2.0%, and 120 min later 6.8% and 1.5%. We recorded differences in infection of duct samples before and after suckling in 40 cases; bacteria were isolated before suckling from six samples, whereas after it from 34 (p < 0.001). Also, we recorded differences in samples collected after suckling and 120 min later in 12 cases; bacteria were isolated immediately post-suckling from eight samples, whereas 120 min later from four (p = 0.375). No significant changes were seen for secretion. We found neither difference between ewes with single or twin lambs, nor among stages of lactation. Mostly, we isolated staphylococci: 70% of isolates before suckling, 80% of isolates after it, 91% of isolates 120 min later. After suckling we also isolated two Mannheimia haemolytica strains. Suckling predisposes to entrance of bacteria into the teat; however, increased teat infections did not result in mammary infections. Isolation of M. haemolytica post-suckling indicates that lambs act as source of infection for this pathogen.     

Update on the development of a novel dry cow therapy using a bismuth-based intramammary teat seal in combination with the bacteriocin lacticin 3147

: Public concerns over the widespread prophylactic use of antibiotics have led to a search for alternatives to dry cow therapy for the prevention of intramammary infections. A popular alternative is to infuse a teat seal at drying-off. The teat seal is a viscous non-antibiotic formulation and when it is infused into the teat canal and the teat sinus it forms an internal seal that provides a physical barrier to invasion by mastitis-causing pathogens. Enhancement of teat seal formulations may be achieved using non-antibiotic additives such as bacteriocins, potent proteins produced by some bacteria that have the ability to kill other microorganisms. This paper traces the history of investigations at Moorepark Research Centre into the efficacy of teat seal plus lacticin 3147, a bacteriocin produced by Lactococcus lactis DPC3147, in the prevention of intramammary infections in dry cows. Indications from on-going investigations are that a dry cow formulation combining the two products has considerable potential as a non-antibiotic prophylactic product.     

Electron microscopic study of leucocytic infiltration of the mammary teat duct during infection with Staphylococcus aureus

Leucocytic response to Staphylococcus aureus infection was observed in the bovine mammary teat duct using transmission electron microscopy. Leucocytes migrating across the stratified squamous epithelium were observed in close association with areas colonised by cocci. Leucocytes gained access across the epithelium to the teat duct lumen by: passage as luminal cells desquamated; migration through degenerate cell cytoplasm; and penetration of cell junctions. The results provide evidence of marked leucocytic infiltration into ductal tissue which may participate in the cellular response to mastitis.     

Detection of contagious ovine ecthyma (orf) and risk factors for infection in small ruminants in Iran

The primary cause of contagious ecthyma is the orf virus, the parapoxvirus prototype. It is a viral problem observed in goat and sheep flocks in Iran, causing economic loss. Orf is a zoonosis with little epidemiological investigation present in Iran. The current research aims at determining the status of this virus, and a PCR was used as a confirmatory instrument. We sampled 668 goats and sheep and various breeding systems. Besides, the orf prevalence was studied, and vaccination efficacy was determined. Moreover, the potential risk factors surveyed for infection with ecthyma were identified. Samples were taken from goat and sheep flocks in the present cross-sectional research, and PCR was used for testing orf DNA. A checklist including animals’ general information was completed. Data were analyzed using univariate tests (chi-square and t-tests) and multivariable binary logistic regression analysis. Three hundred one (45%) goats and sheep detected orf DNA. The age of 70% of positive cases was below one month. Ecthyma infection was significantly higher in imported breeds (87.3%) than indigenous (39.3%). Ninety-six percent of infected goats and sheep in the present work were not vaccinated against ecthyma. The high prevalence of the orf virus was confirmed among goat and sheep flocks in Iran. It is necessary to train ranchers regarding sanitary actions, quarantine, and application of orf vaccination plans.     

Effect of vacuum level on milking efficiency, somatic cell counts (SCC) and teat end wall thickness in ewes of Greek mountain Boutsiko breed

The aim of this work was to determine the effect of vacuum level of machine milking on milk yield, milk composition, milking efficiency, somatic cells counts (SCC), and teat end thickness of the Greek mountain breed Boutsiko ewes. Seventy two ewes after a suckling period of 45 ± 5 days were divided into three equal groups of 24 animals for level of milk production, prolificacy and lactation number. The overall experiment lasted 14 weeks. All groups were milked with a pulsation rate of 120/min, pulsator ratio of 50/50, and three vacuum levels of 38, 44 and 50 kPa, respectively, were applied for the three groups. Every two weeks, in a.m. and p.m. milking, measurements were taken for machine milk (MM) and machine stripped milk (MSM), milk composition, SCC, milk emission characteristics (only in a.m. milking) and teat end wall thickness (TEWT) before and after milking. The results have shown that the lower vacuum level of 38 kPa increased (P < 0.05) the percentage of machine milk (82.9%). There were no significant differences among the groups for milk composition, milk flow rate and SCC, but there was a difference in log SCC (P < 0.05) between 38 and 50 kPa. The teat end wall thickness was lower at the lower vacuum level (38 kPa) after milking, and increased (P < 0.001) as the vacuum level was raised to 50 kPa. The findings of this study suggest that with a lower vacuum level of 38 kPa, the ewes are milked better than those with 44 or 50 kPa. Also, the irritation of teats is considerably reduced, while no differences were seen regarding the health of the udder.     

Investigation of the association between the interval from internal teat sealant administration to calving and early lactation clinical mastitis in New Zealand dairy heifers

AIMS: To examine the association between the interval from internal teat sealant (ITS) administration to calving and the incidence of farmer-recorded clinical mastitis in the first 30 days of lactation in pasture-based dairy heifers.

METHODS: Heifers that were administered an ITS by a single veterinary business in the South Island of New Zealand over the winter of 2014 were enrolled in a cross-sectional observational study. ITS was administered to all heifers on each participating farm on a single calendar day. The dates of calving and farmer-diagnosed clinical mastitis were recorded by farm staff. The interval from ITS administration to calving was categorised into four approximately evenly sized groups: <35, 35–48, 49–69 and >69 days. The quartile of the farm’s calving period in which each heifer calved was also investigated as a potential confounding variable. A hierarchical logistic regression model was constructed to determine the association between the interval from ITS administration to calving with the odds of clinical mastitis in the first 30 days of lactation.

RESULTS: Analysis was performed on 7,126 eligible heifers from 31 farms, with ITS administered between 9 May and 11 July 2014. The mean interval from ITS administration to calving was 52.9 (SD 24.4, min 1, max 137) days. Clinical mastitis was diagnosed in 420/7,126 (5.9 (95% CI=5.4–6.5)%) heifers between calving and day 30 of lactation. In the final multivariable model, which included calving period quartile, interval from ITS administration to calving was not associated with the odds of clinical mastitis (p=0.516). Compared to an interval from ITS administration to calving of <35 days, the adjusted OR of clinical mastitis for intervals of 35–48, 49–69 and >69 days were 0.83 (95% CI=0.59–1.17), 0.71 (95% CI=0.45–1.11) and 0.68 (95% CI=0.36–1.29), respectively.

CONCLUSIONS AND CLINICAL RELEVANCE: Within the range of intervals from ITS administration to calving observed in this study, there was no association with the odds of clinical mastitis in the first 30 days of lactation in dairy heifers. This study suggests that veterinary clinics may be able to extend their ITS administration service and treat dairy heifers earlier than the current recommendation of approximately 4 weeks before the planned start of calving.     

Clinical and ultrasonographic findings in the mammary gland of sheep

AIM: To compare clinical and ultrasonographic findings and assess the value of ultrasonography for evaluating the structure and condition of the teat and teat canal of sheep.

METHODS: The mammary glands of 30 lactating ewes were examined clinically and using ultrasonography. The distance from the teat tip to the ground was measured and diagnostic images of the body of the gland, gland cistern and teat cistern were obtained using an 8.5 MHz frequency ultrasonic transducer. A California mastitis test (CMT) and bacteriological culture were performed on milk samples collected from each half of each udder.

RESULTS: Udder parenchyma and teats were palpably normal in 27/30 (90%) ewes. Milk clots or abnormalities were evident in 4/30 (13%) udders, lesions in the glandular parenchyma and/or teats were evident by palpation in 3/30 (10%), and pathogenic bacteria were cultured from 2/30 (7%). In both of the latter cases, mastitis was diagnosed based on clotted and discoloured appearance of milk and highly positive CMT reactions. Ultrasonographic examination in these udders revealed non-homogenous regions in the glandular tissue and alternating areas of hyperechogenic and hypoechogenic tissue. Overall, highly positive CMT reactions were evident in milk from 14/30 (47%) left halves and 9/30 (30%) right halves (p=0.02). No relationship between teat-to-ground distances and CMT results was evident (p>0.05). Teat canal lengths ranged from 5.7–10.3 mm (mean=8.6; SD=1.3 mm) and the widths from 1.8–3.1 mm (mean=2.3; SD=0.4 mm). Milk sampled from ewes that had long (≥9 mm) or wide (≥2 mm) teat canals was highly positive to the CMT in 75% and 83% of cases, respectively.

CONCLUSIONS: Ultrasonographic measurements of the length and width of the teat canal appeared to correlate with CMT results. The use of ultrasonography in evaluating the health of the udder of small ruminants holds promise for the future.     

Effect of Staphylococcus aureus and Streptococcus uberis on apoptosis of bovine mammary gland lymphocytes

The aim of this study was to determine whether lymphocyte apoptosis is modulated by infections caused by Staphylococcus aureus and Streptococcus uberis. Samples of cell populations were obtained by lavage of the mammary glands at 4 intervals (24, 48, 72 and 168 h) following infection. The percentage of apoptotic lymphocytes peaked at 168 h after challenge with S. aureus or S. uberis. Subsequent experiments focused on in vitro cultivation of mammary gland lymphocytes with S. aureus and S. uberis. These experiments showed a lower percentage of apoptotic lymphocytes following 3 h of cultivating cells with bacteria than after cultivation without bacteria. The results demonstrate that during both experimental infection of bovine mammary glands with S. aureus or S. uberis and during in vitro cultivation of lymphocytes with S. aureus or S. uberis, apoptosis of lymphocytes is delayed.     

The bacterial flora in the teat duct of ewes can protect against and can cause mastitis

We studied the possible effects of bacterial populations within the teat duct, in the pathogenesis of ovine mastitis. In experiment I, 32 ewes were allocated into group A (ewes from which we isolated (+++ growth) coagulase-negative staphylococci), B (ewes from whose duct we isolated (+ growth) coagulase-negative staphylococci) or C (ewes from which we isolated Bacillus spp.) and subdivided into A1, B1, C1 (n=4; challenged by deposition of 1.250 cfu of Mannheimia haemolytica into the teat duct) or A2, B2, C2 (n=4; used as uninoculated controls); group D (n=8) contained ewes with no bacteria in their teat ducts and were challenged as above. There were less bacteriological isolations of flora (P = 0.018) and challenge (P<0.05) organisms from A1 than from A2 and D ewes; the severity of pathological findings in A1 (summed up score: 27) ewes was smaller than in D (summed up score: 36) ewes (P = 0.038). No such findings were evident with B1 or C1 ewes (P>0.4). In experiment II, ewes (groups E and F, n=6) from whose duct we isolated coagulase-negative staphylococci (+ growth) were used; in group G (n=6) ewes with no bacteria in their teat ducts were included. Teat chapping was applied in E and G ewes. All E ewes developed acute clinical mastitis within 24 h after teat chapping, although we had carried out no challenge; there were more bacteriological isolations of flora organisms from E than from F and G ewes (P < 0.001); the severity of pathological findings in E (score: 28) was greater than in F (score: 3) or G (score: 14) ewes. In experiment III, eight ewes with no bacteria in their teat ducts were allocated into group H or I (n=4) and challenged into the teat (group H) or into the gland (group I) with 10(6) cfu of a Staphylococcus simulans recovered from the teat duct of a group E ewe. Group H ewes developed transiently clinical followed by subclinical mastitis (based on bacteriological and cytological evidence), whilst group I ewes developed severe clinical disease. We conclude that staphylococcal flora present in high numbers within the teat duct of ewes can afford some protection against invading microorganisms. However with impeded defence mechanisms of the teat, the same flora may invade the mammary parenchyma and cause clinical mastitis.     

The cytokines of bovine mammary gland: prospects for diagnosis and therapy

The lack of efficacy of conventional strategies for the maintenance of healthy udders in domestic cattle has prompted studies on the use of cytokines for this purpose. The adjuvant use of recombinant bovine cytokines, such as IL-2, IFN-gamma and TNF-alpha, in normal mammary gland, mobilizes innate and acquired immunity. However, stimulated immunity does not prevent or eradicate infection, particularly in the case of Staphylococcus aureus mastitis. Cytokines do, however, improve the bactericidal efficiency of certain antibiotics. The subtle and sensitive changes in the cytokine network of normal and mastitic bovine mammary gland may encourage the use of cytokines in the diagnosis and prognosis of udder health. Numerous studies support this hypothesis, and detection and monitoring of cytokines could become an important alternative management for udder health. The use of cytokines in the immunotherapy, diagnosis and prognosis of mastitis will grow with knowledge of the cytokine network in bovine mammary glands and the development of efficient cytokine diagnostic techniques.     

Incidence risk and aetiology of mammary abnormalities in dry ewes in 10 flocks in Southern Greece

In a field investigation of 10 flocks in Southern Greece, 3367 dairy ewes were examined twice, in order to estimate the incidence risk and the aetiology of mammary abnormalities during the dry-period. Abnormal secretion, lumps, nodules, diffuse hardness, abscesses and cysts were the abnormalities detected. The cumulative incidence of mammary abnormalities during the dry-period was 5.1% (95% confidence interval: 4.4–5.8%); 47% of the cases detected developed during the first three weeks after cessation of lactation. Despite variation in the flock size, there was no between-flock variation in the risk of a ewe developing mammary abnormalities. Staphylococci (Staphylococcus aureus and coagulase-negative isolates) were the most frequently isolated bacteria from mammary samples; Actinomyces pyogenes, Clostridium perfringens, streptococci and Escherichia coli were also isolated. Resistance was encountered among the staphylococcal isolates.     

Seroprevalence and associated risk factors of Chlamydia abortus infection in ewes in Tunisia

Enzootic abortion of ewes (EAE) caused by Chlamydia abortus is a disease of ruminants that results in serious economic losses in livestock industry. The zoonotic potential of the pathogen adds a public health concern on the efforts to control the disease. We report herein a cross-sectional study that was conducted during the lambing season (June and July) in Tunisia to estimate the seroprevalence of C. abortus infection in large sheep herds with abortion history. A total of 803 ewes were sampled and tested using indirect enzyme linked immunosorbent assay (iELISA). The overall apparent seroprevalence at herd and individual levels were 58 % (95 %CI = 39–74.5 %) and 6.6 % (95 %CI = 4.9–8.3 %), respectively. Significant risk factors investigated using univariate and multivariate analyses were history of infertility (OR = 5.7; 95 %CI = 3.05–10.66), the number of reproductive ewes (OR = 2.1; 95 %CI = 1.12–3.94), the control of new animals at introduction (OR = 4.35; 95 %CI = 2.46–7.68), the sharing of drinking water (OR = 2.18; 95 %CI = 1.22–3.9), the exchange of breeding males (OR = 2.56; 95 %CI = 1.003–6.54), the disposal of abortion materials without precaution (OR = 4.36; 95 %CI = 2.42–7.87), the lack of lambing barn (OR = 2.39; 95 %CI = 1.13–5.04), the non-application of hygienic post-abortion measures (OR = 10.35; 95 %CI = 5.28–20.26) and the manure management (OR = 11.35; 95 %CI = 3.26–39.48). To the best of our knowledge, this is the first sero-epidemiological survey conducted on an abortive disease in Tunisian ewes that investigated the risk factors of C. abortus infection.     

Pregnancy and lactation affect the microvasculature of the mammary gland in mice.

Microvascular changes in the mammary gland in mice during pregnancy and lactation were investigated by scanning electron microscopy (SEM) with a corrosion cast method, transmission electron microscopy (TEM) and morphometry. By SEM, duct-associated capillary plexuses were sparsely distributed to branch into adipocytes during virgin period. With advance in pregnancy, both branches from the capillary plexuses and branches from the vessels surrounding adipocytes extended further to form capillary networks. The basket-like architecture was completed by day 18 of pregnancy. These findings may indicate that angiogenesis occurs frequently during this period. During lactation, the basket-like architecture still remained and the capillaries surrounding alveoli meandered. After weaning, the regression of microvasculature followed the degeneration of alveoli. By TEM and morphometry, the density of pinocytotic vesicles (PVs) (number of PVs per microns 2 of endothelium cytoplasm) increased twofold from day 18 of pregnancy to day 5 of lactation, furthermore increased threefold from days 10 to 20 of lactation, and subsequently decreased after weaning. Marginal folds and microvillous processes gradually increased in length with advance in pregnancy, reached the maximum from days 5 to 15 of lactation, and thereafter decreased. In addition, the capillaries with thinner walls were in close contact with alveoli during the late stage of pregnancy and during lactation. Furthermore, the alveolar epithelial cells had well-developed basal infoldings during lactation. These findings suggest that the capillaries play an important role in transporting materials necessary for milk production.