Antimicrobial susceptibility of Prototheca zopfii isolated from bovine intramammary infections

In vitro antimicrobial susceptibility tests were carried out on 48 strains of Prototheca zopfii, an achlorophyllous algae causing refractory mastitis in dairy cows; 27 antimicrobials were evaluated. All strains were susceptible to both myxin and nystatin. In addition, 22 strains were susceptible to amphotericin B, 21 to polymyxin B, and 18 to gentamicin. Only 1 strain was susceptible to kanamycin. All strains were resistant to ampicillin, bacitracin, carbenicillin, cephalothin, chloramphenicol, clotrimazole, cloxacillin, erythromycin, flucytosine, ketoconazole, lincomycin, miconazole, neomycin, nitrofurazone, novobiocin, oleandomycin, penicillin, rifampin, streptomycin, tetracycline, and vancomycin.     

Eradication of Prototheca zopfii infection in a dairy cattle herd

Protothecosis is a severe form of mastitis in dairy cows caused by colorless algae of the genus Prototheca. Since P. zopfii is highly resistant to all known chemotherapeutics, infected cows must be removed from the herd. Eradication measures are difficult since many chronically infected cows may become intermittent shedders. Therefore, cultural methods are insufficient for control measures. In order to eradicate Prototheca zopfii-mastitis in dairy cattle herds, two isotype specific indirect ELISA for detection of IgA and IgG1 in whey were used in a dairy herd highly affected with protothecal mastitis. All cows (n = 313) were tested four times in intervals of six months. Milk specimens were examined in parallel by cultivation and serologically using two indirect ELISA systems for specific IgA and IgG1 in whey. Cows tested Prototheca positive were consequently separated from the herd and slaughtered. At the first examination, 15.6% of the animals were found positive by culture, and 23.3% were positive in at least one of the ELISA systems. Within two years, protothecal prevalence and incidence decreased to zero indicating that the eradication strategy used was successful. In summary, serological identification of P. zopfii-infected lactating cows is an useful tool to eradicate protothecal bovine mastitis in infected herds.     

Prototheca zopfii genotypes isolated from cow barns and bovine mastitis in Japan

This study is the first investigation on Japanese isolates of Prototheca zopfii from bovine mastitis and the cow-barn surroundings by molecular characterization to clarify routes of infection for bovine protothecal mastitis. We performed isolation of Prototheca from cow-barn surroundings (drinking water, sewage and feces) and milk samples from cases of bovine mastitis. Genotypes of the 32 isolates of P. zopfii from cow-barn surroundings and 67 isolates from mastitis were analyzed by genotype-specific PCR assays and restriction fragment length polymorphism (RFLP) assays. All mastitis isolates were identified as P. zopfii genotype 2. Conversely, 29 isolates from cow-barn surroundings were identified as P. zopfii genotypes 1 and 3 isolates as genotype 2, respectively. Given these results, both genotypes of P. zopfii could exist in cow-barn surroundings, but no sites were identified as frequent sources of P. zopfii genotype 2. P. zopfii isolates should thus be further explored with regard to genotype to clarify the reservoir of etiological agents in bovine Prototheca mastitis.     

奶牛中型无绿藻性乳房炎

目前已有12个国家报道无绿藻性奶牛乳房炎,造成严重的经济损失并对公共安全存在潜在的危害.2010年春季,北京地区某奶牛场暴发中型无绿藻引起的临床型乳房炎.从患有临床型乳房炎的10头奶牛的11个乳区分离到8株酵母样微生物,鉴定为中型无绿藻.用16种抗生素药敏纸片进行药敏试验,结果显示中型无绿藻仅对庆大霉素、链霉素和阿米卡星等氨基糖苷类抗生素敏感.同化试验证明该分离株能发酵葡萄糖和丙三醇.本试验在我国首次报道了中型无绿藻能引起奶牛临床型乳房炎.     

Paraparesis as initial manifestation of a Prototheca zopfii infection in a dog

A case of protothecosis causing non‐ambulatory paraparesis in a dog without clinical evidence of disseminated infection is described. A five‐year‐old female Labrador retriever was referred with a 10‐day history of progressive non‐ambulatory paraparesis and lumbar pain as the only physical and neurological abnormalities. Lumbar myelography revealed severe extradural spinal cord compression extending from L4 to L7 vertebrae, and a right hemilaminectomy was performed. Surgical findings included an adherent whitish hard ill‐defined mass. Cytology and biopsy results disclosed the presence of algae enclosed in a matrix of chronic inflammatory infiltrate. Culture confirmed the presence of Prototheca species. Neurological improvement occurred within a month, and the dog received antifungal treatment without evidence of clinical disseminated disease for 6 months, but died after a generalised tonic–clonic seizure. Post‐mortem examination revealed multiple foci of inflammatory granulomatous infiltrate and algae‐like structures in the brain, lumbar intumescence and cauda equina. Prototheca zopfii was identified using molecular biology methods.     

Stochastic bio-economic model of bovine intramammary infection

Although the dynamics of transmission play an important role in the occurrence of intramammary infection (IMI), they have not been considered in previous models used to estimate the cost of IMI. The bio-economic model described includes within-herd dynamics of pathogen-specific IMI. The model simulated Staphylococcus aureus, Streptococcus uberis, Streptococcus dysgalactiae, and Escherichia coli IMI stochastically and estimated the cost of these IMI in a herd of 100 dairy cows in a situation where a quota is applied to milk production. A Reed–Frost model was used for S. aureus, S. uberis, and S. dysgalactiae IMI and a Greenwood model for E. coli IMI. Economic analysis was conducted per pathogen for clinical and subclinical IMI. The parameters used in the model were based on the literature and were deemed credible and valid. Median annual incidence of clinical and subclinical IMI for all pathogens varied considerably. This variation was greatest for S. aureus IMI. The annual incidence of IMI in a herd of 100 dairy cows caused by S. aureus varied between 0 and 88 cases, with a median of 5 cases and the 5th and 95th percentiles of 0 to 36 for clinical IMI, and a median of 7 cases with the 5th and 95th percentiles of 0 to 52 for subclinical IMI. In consequence, the average total annual net costs also varied widely for S. aureus IMI. Clinical IMI costs were € 1375, with the 5th and 95th percentiles of 0 to 4716 and subclinical IMI costs were € 1219, with the 5th and 95th percentiles of 0 to 4030. The average annual net cost due to the 4 simulated pathogens combined was € 4896 and varied from € 915 to € 11,287 in a herd of 100 dairy cows. The bio-economic model developed for this study will be utilized as a tool to investigate the economic impact of management of pathogen-specific IMI.     

Prototheca zopfii mastitis in dairy herds under continental climatic conditions

In the last 2 years 223 cases of bovine mastitis caused by Prototheca zopfii infection were identified in 32 large-scale dairy herds. All of these farms were in Hungary, which has a continental type, temperate zone climate. Both the sporadic and epidemic forms of P. zopfii mastitis were observed. All the herds affected by the epidemic form had poor hygienic conditions and suffered from several managerial faults, but no specific predisposing factors could be identified. In almost all of the cases, the type II variant of this pathogen was isolated; however, the type III variant was isolated from three cows. The cows had a higher chance of new infection in the early weeks of lactation and in the summer. The P. zopfii infection usually resulted in a chronic subclinical, or mild clinical, inflammatory process in the udder, and was followed by a dramatic loss in milk production and a permanent increase in somatic cell count. The histopathological findings could be characterized as a progressive interstitial mastitis associated with alveolar atrophy. The self-recovery rate was very low.     

Prototheca zopfii mastitis in a herd of dairy cows

Mastitis caused by the colourless alga Prototheca zopfii was diagnosed in 17 of 120 cows in a dairy herd. Infection occurred in animals varying from 3-14 years old and was present in one to four quarters of each cow. Nine cases were associated with clinical mastitis characterised by the presence in milk of flakes or small clots. Somatic cell counts consistent with subclinical mastitis (>500 x 10(3) cells/ml) were recorded in five of the eight remaining cows. Histological examination of udder tissue showed the presence of granulomatous lesions associated with the presence of Prototheca. The problem was identified and controlled by repeated microbiological examination of milk samples from all lactating cows and immediate culling of infected animals. P. zopfii was also recovered from environmental water samples on this farm. It is suggested that infection may have occurred as a result of teat sores caused by trauma from a milking machine, and the tendency for cows to lay down on a race, the surface of which was sometimes flooded by drain water in which Prototheca were present.     

Prototheca zopfii mastitis in a herd of dairy cows

Mastitis caused by the colourless alga Prototheca zopfii was diagnosed in 17 of 120 cows in a dairy herd. Infection occurred in animals varying from 3–14 years old and was present in one to four quarters of each cow.

Nine cases were associated with clinical mastitis characterised by the presence in milk of flakes or small clots. Somatic cell counts consistent with subclinical mastitis (>500 × 10³ cells/ml) were recorded in five of the eight remaining cows.

Histological examination of udder tissue showed the presence of granulomatous lesions associated with the presence of Prototheca.

The problem was identified and controlled by repeated microbiological examination of milk samples from all lactating cows and immediate culling of infected animals.

P. zopfii was also recovered from environmental water samples on this farm. It is suggested that infection may have occurred as a result of teat sores caused by trauma from a milking machine, and the tendency for cows to lay down on a race, the surface of which was sometimes flooded by drain water in which Prototheca were present.     

Review of the microbiological, pathological, and clinical aspects of bovine mastitis caused by the alga Prototheca zopfii

The mastitis caused by P. zopfii alga is a disease of high-producing, machine-milked dairy cows. It occurs worldwide in tropical and temperate climatic areas, and mostly appears sporadically in a therapy-resistant form. However, in poorly managed dairy herds it may be endemic, causing serious economic losses as a result of decreased milk quality and quantity and culling of infected animals. The biological properties of this pathogenic alga, the laboratory methods available for its isolation and identification, the pathological and clinical features of this form of mastitis, and the principles of its control are reviewed in this paper.     

Bovine mastitis caused by Prototheca zopfii: a case study

An outbreak of severe indurative mastitis in newly calved cows, from which Prototheca zopfii was isolated, is described. The refractory nature of this type of mastitis and the ineffectiveness of the presently available intramammary preparations are highlighted.     

Occurrence of Prototheca zopfii, a mastitis pathogen, in milk

Prototheca zopfii was isolated from cow-composite milk and bulk tank milk by a new selective Prototheca enrichment method. Repeated testing of cow-composite milk from individual cows resulted in P. zopfii isolation data indicating a strong statistical correlation of P. zopfii with specific cows. Prototheca sp. were isolated from the milk of 31 of 79 cows in a single herd, and contamination was discounted as the source. Prototheca sp. were also recovered from 28 of 787 bulk tank milk samples from the Eastern U.S.A., and 22 of 69 temperature control milk samples from pooled dairy farm milk from delivery trucks.     

In vitro algaecide effect of disinfectants on Prototheca zopfii genotypes 1 and 2

Bovine mastitis due to Prototheca zopfii leads to reduced milk production and is difficult to cure. Therefore, prevention is the best approach and this is best achieved through the use of effective disinfectants. The aim of this study was to evaluate the in vitro algaecide efficacy of conventional disinfectants against strains of P. zopfii genotype 1 and 2. The minimal algaecide concentration (MAC) of alkyldiaminoethylglycine hydrochloride, chlorhexidine, dioxide chlorine, povidone iodine and sodium hypochlorous acid against 10 isolates and the type strain (SAG2063(T)) of P. zopfii genotype 1 as well as 10 isolates and the type strain (SAG2021(T)) of P. zopfii genotype 2 were examined using the micro dilution method. This in vitro study indicated that alkyldiaminoethylglycine hydrochloride, chlorhexidine, povidone iodine and sodium hypochlorous acid, but not dioxide chlorine, are effective against both genotypes of P. zopfii.     

Natural infection of red deer with bovine tuberculosis

Six bovine tuberculosis-free red deer hinds were introduced in October 1993 to a 1.8 ha enclosure, within a larger field study site known to contain tuberculous possums, and kept there for 9 months. A Mycobacterium bovis-infected possum was found in the vicinity of the deer enclosure 3 weeks after the introduction. Subsequently, a further eleven infected possums were found in the area. The deer were monitored by repeated composite antibody detection ELISA and lymphocyte transformation assays for tuberculosis, interpreted in parallel, by skin testing and by routine culturing of samples collected from potential excretion sites. Lymphocyte transformation assay evidence of M. bovis infection in four hinds was first observed 4 months after introduction. One other hind became bovine tuberculin lymphocyte transformation assay positive in the 5th month. Positive or equivocal bovine reactivity remained evident at most test episodes. A comparative cervical skin test performed in July 1994, shortly before slaughter, was positive in these five hinds. Mycobacterium bovis was recovered off swabs from the oropharyngeal tonsils of two hinds during routine sampling. Detailed necropsy of the six deer revealed a single typical tuberculous lesion in only one, but culturing of various tissue specimens ascertained that the five blood test and comparative cervical skin test-positive animals were all infected. Mycobacterium bovis was cultured from the oropharyngeal tonsils of four and medial retropharyngeal lymph nodes of two of the deer with no typical gross lesions. Six additional tuberculosis-free hinds were introduced to the enclosure in April 1994 and kept there for 12 months. Four of these animals showed a positive lymphocyte transformation assay response to M. bovis after 9 weeks, but no significant reactivity thereafter. Concurrent observational studies suggest that five of the first six deer probably became infected through close inspection and investigation of the tuberculous possums, although the possibility of deer-to-deer transmission cannot be totally excluded. The likely deer-possum contact, and thus exposure to M. bovis, was related to the curiosity and social ranking of the hinds. The second group appear to have had transient exposure to M. bovis, possibly caused by direct contact with the infected hinds introduced earlier. This group never showed any curiosity toward, or interaction with, possums during the periods of observation.     

Innate immune response in experimentally induced bovine intramammary infection with Staphylococcus simulans and S. epidermidis

ABSTRACT: Coagulase-negative staphylococci (CNS) are in several countries the most common bacteria isolated in subclinical mastitis. To investigate the innate immune response of cows to infections with two common mastitis-causing CNS species, Staphylococcus epidermidis and Staphylococcus simulans, experimental intramammary infection was induced in eight cows using a crossover design. The milk somatic cell count (SCC), N-acetyl-β-D-glucosaminidase (NAGase) activity, milk amyloid A (MAA), serum amyloid A (SAA) and proinflammatory cytokines interleukin (IL)-1β, IL-8, and tumor necrosis factor α (TNF-α) were determined at several time points before and after challenge. All cows became infected and showed mild to moderate clinical signs of mastitis. The spontaneous elimination rate of the 16 infections was 31.3%, with no difference between species. Infections triggered a local cytokine response in the experimental udder quarters, but cytokines were not detected in the uninfected control quarters or in systemic circulation. The innate local immune response for S. simulans was slightly stronger, with significantly higher concentrations of IL-1β and IL-8. The IL-8 response could be divided into early, delayed, or combined types of response. The CNS species or persistency of infection was not associated with the type of IL-8 response. No significant differences were seen between spontaneously eliminated or persistent infections.     

Observations on intramammary infection and somatic cell counts in cows treated with recombinant bovine somatotropin.

Data were collected on udder health variables as part of a study of the effects of recombinant bovine somatotropin on production in lactating dairy cows. Milk samples, obtained at three intervals during the study, were assessed for their somatic cell count and bacteriological culture result. There was an increase in the prevalence of infection at mid-lactation in the 20.6 and 41.2 mg per day treatment groups as compared to the controls. There was no difference detected in the mean cell count between groups from the samples collected pretrial, mid-lactation, or late lactation. However, analysis of the individual cow Dairy Herd Improvement somatic cell count data showed a difference between groups which was most evident in mid-lactation.     

The use of polyethylene intramammary device in protection of the lactating bovine udder against experimental infection with Staphylococcus aureus or Streptococcus agalactiae.

The susceptibility of lactating bovine udder quarters fitted with a polyethylene intramammary device to infection was investigated. Following experimental challenge with Streptococcus agalactiae or Staphylococcus aureus, the incidence of infection was significantly (p less than 0.05) lower in intramammary device-fitted quarters compared to control quarters. In general, total foremilk and strippings milk somatic cell counts for intramammary device-fitted and control quarters were not significantly (p less than 0.05) different. Differential foremilk and strippings milk somatic cell counts were significantly (p less than 0.05) higher in samples from intramammary device-fitted quarters compared to control quarters.