Presence of subepithelial lymphoid nodules in the teat of ewes

A total of 87 clinically healthy ovine teats were examined bacteriologically (by scraping the mucosa) and histologically. Teats examined were those of lactating mammary glands with no bacteria isolated (n = 23); of mammary glands after cessation of lactation with no bacteria isolated (n = 25); of lactating mammary glands with bacteria isolated (n = 22); and of mammary glands after cessation of lactation with bacteria isolated (n = 17). The salient histological feature was subepithelial leucocytic infiltration. In teat cisterns, lymphocytes were the predominant cell type and in teat ducts, lymphocytes and neutrophils were seen in equal proportions. Subepithelial lymphoid nodules, some with germinal centres, were detected in 43 (49%) teats. The majority of lymphoid nodules was observed at the border between teat duct and teat cistern. Presence of bacteria was significantly associated with the presence of leucocytic activity (P < 0.001) and with the presence of lymphoid nodules (P = 0.032). We conclude that the presence of induced subepithelial lymphoid tissue at the border between teat duct and teat cistern appears to be important in protecting the mammary gland during the early stages of bacterial invasion. The findings call for further investigations into the lymphoid structures of the teat; these should elucidate the role and development of mammary mucosa-associated lymphoid tissues and may lead to strategies for enhancing non-specific defence mechanisms of the mammary gland.     

The influence of anti-inflammatory therapy on bacterial clearance following intramammary Escherichia coli challenge in goats

Coliform mastitis in dairy cattle frequently results in systemic disease with occasional deaths in association with endotoxic shock. Systemic anti-inflammatory therapy has been used to alter the course of endotoxic shock in severe cases. Use of anti-inflammatory therapy has been questioned on the basis that such treatment may compromise immune function and decrease clearance of bacteria from infected mammary glands. The purpose of this investigation was to determine whether anti-inflammatory therapy influenced bacterial clearance following intramammary challenge of lactating goats with Escherichia coli.Standardized quantities of a pathogenic coliform culture were infused through the teat canal into one half of the mammary gland in 18 goat does. The does were then randomly assigned to receive one of three intravenous treatments: saline (controls), one dose of steroid (dexamethasone), or two doses of a non-steroidal anti-inflammatory agent (flunixin meglumine). The clinical signs, milk production, complete blood counts, serum clinical chemistry values, milk bacterial cultures and milk somatic cell concentrations were monitored sequentially.Goats treated with anti-inflammatory agents exhibited some improvement in clinical response to challenge with E. coli (e.g. rectal temperature, degree of appetite suppression) as compared to saline controls. There were no significant differences between treatments in the degree of inflammation present in the mammary glands or supramammary lymph nodes examined at necropsy. The most important finding was that anti-inflammatory therapy did not adversely influence the clearance of E. coli from challenged glands.     

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.     

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.     

Outbreak of bovine clinical mastitis caused by Mycoplasma bovis in a North Italian herd

This report describes an outbreak of Mycoplasma bovis mastitis affecting 45 cows in a herd of 122 dairy cattle in Northern Italy. Clinically, the outbreak was characterized by agalactia, multiple swollen and painless quarters, high milk somatic cell count and unresponsiveness to conventional antibiotic therapy. M. bovis was isolated from the milk samples of all the 32 affected cows tested and from the mammary tissue of three affected cows that underwent necropsy. No other pathogens were isolated from these samples. Lesions in two of the necropsied cows were characterized by mild chronic suppurative mastitis and galactophoritis. The other necropsied cow showed a chronic necrosuppurative and pyogranulamaous galactophoritis, a condition not previously associated with M. bovis. M. bovis was detected immunohistochemically in the lumen of the affected mammary ducts suggesting that ascending infection via the teat canal was the likely route of transmission. No other intralesional pathogens were demonstrated microscopically.     

Preparation of an epithelium-free mammary fat pad and subsequent mammogenesis in ewes

The objective of this study was to develop a surgical procedure for the preparation of an epithelium-free mammary fat pad (cleared mammary fat pad; CFP) in ewes. At 7 to 10 d of age, ewe lambs (n = 43, mean BW 9.2 +/- .2 kg at 14 d) were sedated and one mammary gland was locally anesthetized. An incision circumscribing the base of the teat was made and blunt dissection was performed through the extraneous mammary fat pad tissue to enable the parenchyma and teat to be wholly removed. Failure to completely remove the epithelium enabled it to regenerate and grow into the mammary fat pad. Mean diameter of the parenchymal rudiment at 7 to 10 d of age was 8.9 +/- .5 mm (range of 5 to 16 mm). The excision site was closed with wound clips and recovered lambs returned to their dams. The contralateral mammary gland remained intact, allowing it to undergo normal development. Live weight gain was unaffected by this procedure. Ewes were subsequently slaughtered in groups at various stages of prepuberty, puberty, gestation, and lactation. Of 39 operated glands recovered, only one demonstrated epithelial outgrowth within the CFP. Parenchyma within the contralateral, intact gland underwent phases of rapid growth in prepuberty, puberty, and late gestation and was capable of milk synthesis after steroid induction or parturition. Change in weight of the CFP paralleled that of the intact mammary gland to 100 d of pregnancy. Sham CFP surgery was performed on four additional ewes wherein the parenchyma was completely excised and immediately replaced. Sham-operated epithelium populated the mammary fat pad and synthesized milk that could be expressed from the teat. A CFP in sheep will be a useful model for future investigations into the local growth regulatory mechanisms associated with ruminant mammogenesis.     

An increased incidence of mastitis caused by Prototheca species and Nocardia species on a farm in São Paulo,Brazil

Mastitis caused by Prototheca spp or Nocardia spp is considered to be difficult to treat. Both microorganisms are contaminants commonly found in soil. The occurrence of mastitis caused by these agents was studied in a particular dairy farm. In this herd, the animals were kept at pasture overnight and during daytime were brought to a pen where they were fed. This pen accumulated mud and faeces, particularly in the rainy season. During milking, pre-dipping of the teats was performed with an iodide solution, but they were not washed, so a layer of soil and faeces remained which may have contaminated the milking equipment. The herd comprised 91 lactating animals and 47 dry cows. For microbiological examination, 107 milk samples were collected from lactating cows and 186 samples of mammary secretions from the dry cows. Prototheca spp were isolated from 14.55% of the milk samples and Nocardia spp from 4.55%. Prototheca spp were isolated from 8.06% of the secretion samples from dry cows and Nocardia spp were isolated from 2.15% samples. The high occurrence of mastitis due to these environmental agents reflects the problem of keeping animals in muddy pastures and pens, and the defective pre-milking hygiene for the teats.     

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 (> or = 9 mm) or wide (> or = 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.     

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%).     

Pathological and microbiological characterization of mastitis in dairy cows

Mastitis may be caused by a wide range of microorganisms able to induce distinct lesions in mammary tissues. This study aims to characterize the gross and microscopic features of mastitis in dairy cows and to correlate them with the pathogens involved. The udders of slaughtered dairy cows were inspected and milk samples from each mammary quarter or fragments of the parenchyma were sent for microbiological analysis, and tissue collected for histopathological evaluation. A total of 148 cows and 592 mammary quarters were collected. From these, 432 quarters (73%) had mastitis and in 160 (27%), no changes were observed. Mastitis was classified into seven patterns based on the histopathological findings, of which mixed, lymphoplasmacytic, and suppurative mastitides were the most prevalent with 35.9% (155/432), 27.1% (117/432), and 14.3% (62/432) of the cases, respectively. These patterns were associated with the same set of pathogens: Streptococcus spp., coagulase-negative Staphylococcus (CNS), Staphylococcus aureus, Streptococcus agalactiae, Streptococcus uberis, and Corynebacterium bovis. The pyogranulomatous pattern represented 7.2% (31/432) of the cases with distinct distribution based on the agent involved, mostly S. aureus and Nocardia sp. Abscedative mastitis accounted for 6.0% (26/432) of the cases; it was characterized by multiple abscesses in the parenchyma and was mainly caused by Trueperella pyogenes. Necrosuppurative mastitis represented 5.8% (25/432) of the cases which were characterized by severe parenchyma necrosis and were caused by bacteria such as CNS and Escherichia coli. The granulomatous pattern represented 3.7% (16/432) of the cases and was occasionally associated with Mycobacterium sp.

     

Effects of intramammary infusions of interleukin-8 on milk protein composition and induction of acute-phase protein in cows during mammary involution

The effects of interleukin-8 (IL-8) on bovine mammary functions such as milk protein secretion and the blood-milk barrier during mammary involution were evaluated. Following the final milking, recombinant bovine (rb) IL-8 (5 or 25 microg) and a saline placebo were individually infused into the left- and right-front teat cisterns of 6 cows, respectively. Three cows without treatment at the final milking were also used as controls. Mammary secretions and blood were collected at -24, 0, 10, 24, 72, 168, 336, and 720 h after infusion. In the mammary glands infused with 25 microg of rbIL-8, the increases in somatic cell counts and in the concentrations of serum albumin, IgG1 and IgG2, and the decreases in the concentrations of alpha- and beta-casein and beta-lactoglobulin were greater than in the control glands. In the mammary glands infused with 5 microg of rbIL-8, compared to the glands infused with 25 microg of rbIL-8, these changes were moderate. These results indicate that rbIL-8 impairs the integrity of the blood-milk barrier and suppresses milk-specific protein secretions. In the cows infused with 25 microg of rbIL-8, the rectal temperature and serum haptoglobin level were transiently elevated after the infusion, showing that intramammary infusion of rbIL-8 could elicit systemic inflammation.     

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.     

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.     

猪翻乳头的解剖组织学特征的研究与探讨

通过对正常乳头，翻乳头，副乳头及其乳房的组织样本进行解剖学观察，对照，发现正常乳头由乳基，乳头本组成；翻乳头由乳基，不明显的乳头体组成；副乳头只有小突起。对各类乳腺做成切片观察，对照，发现正常乳头和翻乳头均有完整的乳腺，乳管，副乳头的乳腺，乳管不发达，且很稀少。     

Evaluation of three-dimensional ultrasonography of the bovine mammary gland

OBJECTIVE: To evaluate 3-dimensional (3-D) ultrasonography of the bovine mammary gland, particularly of the teat. ANIMALS: 6 lactating cows. PROCEDURES: Clinical and ultrasonographic examinations of mammary glands of each cow were performed. Teats were removed from a slaughtered cow and examined via ultrasonography. All scans were performed by use of a sensorless standard transducer (8.5 to 10 MHz linear array). The 2-dimensional data acquired were downloaded to an off-line system, and software was used to digitize each image and produce a 3-D block of digitized information. The selected anatomic area was displayed as a 3-D volume cube. RESULTS: Good-quality, 3-D views of the entire mammary gland were acquired by obtaining sections through the glandular parenchyma, gland cistern, teat cistern, and teat canal. CONCLUSIONS AND CLINICAL RELEVANCE: 3-D ultrasonography allows perspective images of the mammary gland to be obtained. Many scanners routinely used in veterinary practice have the resolution required to produce satisfactory images, and the cost of refitting a standard ultrasonographic unit with 3-D software is affordable; however, the cost of a volume transducer with a positioning system and mechanical sweep strategy may be prohibitive. Three-dimensional ultrasonography is a new imaging technique that has promising applications in many fields of veterinary medicine.     

Aetiology and histopathology of bovine mastitis of espontaneous occurrence

Infectious bovine mastitis is responsible for serious economic losses in dairy cattle breeding. Information about the damages caused by micro-organisms is of great interest and importance. The purpose of the present study was to report on the microbiological and histopathological aspects of the mammary parenchymas of slaughtered dairy cows. A total of 184 mammary glands were examined and samples of mammary parenchyma were collected for microbiological and histopathological examinations. Micro-organisms were isolated from 69.6% samples; 23 (12.5%) of the 184 samples did not show histological changes; inflammatory response was observed in 56 (30.4%) samples; inflammatory response and repair were present in 82 (44.6%) samples; repair process was verified in 23 (12.5%) of the mammary glands. Coagulase-negative Staphylococci were observed in 53.8% of the samples followed by coagulase-positive Staphylococci (7.6%), Prototheca sp. (2.2%) and Streptococcus dysgalactiae (1.6%). Coagulase-negative Staphylococci and coagulase-positive Staphylococci were associated mainly to chronic inflammatory response and chronic inflammatory response and repair. Samples from which no micro-organisms were isolated (n = 56) had no histological changes in 82.6% of the cases. These results were higher (P < 0.05) when compared to the samples with micro-organisms and without histological changes (17.4%).     

Immunity to Escherichia coli in pigs: Antibody Secretion by the Mammary Gland after Intramammary or Intramuscular Vaccination with an E. coli Vaccine

Indirect hemagglutinating antibody titres in individual gland samples of colostrum and milk from 13 sows were measured. Five of the sows were vaccinated via a mammary gland and five by the intramuscular route with a live formalinised Escherichia coli vaccine and three remained as non-vaccinated controls.

Antibody titres were higher in colostral and milk whey from the vaccinated sows than from non-vaccinated groups. The inoculated gland in the group of sows given vaccine by the intramammary route secreted milk containing markedly more antibodies to the vaccine E. coli strain than did the non-vaccinated glands. Milk from the vaccinated gland did not contain higher titres to heterologous E. coli O antigens than milk from non-vaccinated glands. Serum titres were the same or higher than the titres in colostrum from non-vaccinated glands.

     

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.     

In vitro algaecide effect of sodium hypochlorite and iodine based antiseptics on Prototheca zopfii strains isolated from bovine milk

Prototheca zopfii has been considered one of the most important causes of environmental mastitis in Brazil. These algae are refractory to conventional therapy and cause great damage to the mammary gland. The present study evaluated the in vitro algaecide effect of sodium hypochlorite and iodine based antiseptics on 27 P. zopfii strains isolated from the milk of cattle. Low concentrations of sodium hypochlorite (0.0390625-0.15625%) and iodine (0.15625-0.625%) were effective against the isolates. These antiseptics may be recommended for hygiene routines, pre and postdipping and cauterization of bovine mammary glands infected by P. zopfii.