Etiology and antimicrobial susceptibility of udder pathogens from cases of subclinical mastitis in dairy cows in Sweden

Background

A nationwide survey on the microbial etiology of cases of subclinical mastitis in dairy cows was carried out on dairy farms in Sweden. The aim was to investigate the microbial panorama and the occurrence of antimicrobial resistance. Moreover, differences between newly infected cows and chronically infected cows were investigated.

Methods

In total, 583 quarter milk samples were collected from 583 dairy cows at 226 dairy farms from February 2008 to February 2009. The quarter milk samples were bacteriological investigated and scored using the California Mastitis Test. Staphylococci were tested for betalactamase production and presence of resistance was evaluated in all specific udder pathogens. Differences between newly infected cows and chronically infected cows were statistically investigated using logistic regression analysis.

Results

The most common isolates of 590 bacteriological diagnoses were Staphylococcus (S) aureus (19%) and coagulase-negative staphylococci (CNS; 16%) followed by Streptococcus (Str) dysgalactiae (9%), Str. uberis (8%), Escherichia (E.) coli (2.9%), and Streptococcus spp. (1.9%). Samples with no growth or contamination constituted 22% and 18% of the diagnoses, respectively. The distribution of the most commonly isolated bacteria considering only bacteriological positive samples were: S. aureus - 31%, CNS - 27%, Str. dysgalactiae - 15%, Str. uberis - 14%, E. coli - 4.8%, and Streptococcus spp. - 3.1%. There was an increased risk of finding S. aureus, Str. uberis or Str. dysgalactiae in milk samples from chronically infected cows compared to findings in milk samples from newly infected cows. Four percent of the S. aureus isolates and 35% of the CNS isolates were resistant to penicillin G. Overall, resistance to other antimicrobials than penicillin G was uncommon.

Conclusions

Staphylococcus aureus and CNS were the most frequently isolated pathogens and resistance to antimicrobials was rare.     

Performance of second-season grazing cattle following different levels of parasite control in their first grazing season

A 3-year grazing trial was performed during 2003-2005 on a commercial steer-producing farm in Sweden to study performance of second-season grazing (SSG) cattle following different levels of parasite exposure during their first grazing season. Initially, groups of 10 first-season grazing (FSG) cattle were each year assigned to four parasite control strategies: (1) turn-out onto pasture that during the previous year was grazed by SSG cattle, followed by a mid-July move to aftermath, (2) supplementation with concentrate and roughage for 4 weeks after turn-out, (3) no treatment, or (4) anthelmintic treatment (injectable doramectin) every fourth week. All animals were set stocked, except for those in group one. Next spring and following housing the cattle were turned out for their second grazing season onto approximately 25 ha communal pasturelands as a common mob. Weighing, faecal sampling and blood collection were performed at turn-out and then every 4 weeks for the 20-week grazing season. Faecal samples were also collected on day 10 after turn-out for detection of coccidian oocysts. Antibodies to Dictyocaulus viviparus were analysed at the time of their second housing period, and when elevated levels were recorded, stored serum samples from seropositive animals were analysed retrospectively. Results showed early-season weight losses of up to 47 kg in the SSG cattle. However, faecal egg counts were generally low and there was no correlation between SSG performance and treatment history as FSG cattle. Still, cumulative egg counts were significantly higher in animals that had been treated with anthelmintic as FSG cattle but serum pepsinogen concentrations showed no significant differences and the output of Eimeria alabamensis oocysts seldom exceeded 10,000 oocysts per gram faeces. Antibodies to D. viviparus were observed from July 2004 and from June 2005 but not in 2003. In 2004 and 2005, 64% and 83% of the animals, respectively, were seropositive for D. viviparus. It is concluded that weight gain penalties resulting from different levels of parasite infections the first grazing season remained during the second grazing season in 2004 and 2005 but no differences in weight gain could be identified in SSG cattle that had experienced different parasite control measures during their first grazing season. This was under conditions where the level of pasture infectivity was low to moderate during the second grazing period. However, D. viviparus was demonstrated in SSG animals during the two last years of the study and was an important confounding factor.     

Expulsion of small strongyle nematodes (cyathostomin spp) following deworming of horses on a stud farm in Sweden

This study was conducted on a stud farm in Sweden to investigate the species composition of cyathostomins expelled in the faeces of horses after deworming using three different anthelmintic preparations. Twenty-seven horses excreting > or = 200 strongyle eggs per gram faeces (EPG) were divided into three comparable groups and dewormed on day 0 with either of following compounds: 0.2 mg ivermectin per kg body weight (bw), 19 mg pyrantel pamoate per kg bw or 7.5 mg fenbendazole per kg bw. For each of the 3 days following anthelmintic treatment faeces was collected from individual horses and subsamples were fixed in formalin. Four days after the anthelmintic treatment all horses were re-treated with ivermectin and faeces was collected on day 5. Individual subsamples from each of the four sampling occasions were examined for cyathostomin nematodes. Sixty-three to 270 worms per horse were identified to the species level. The majority of the worms recovered were expelled during the first day from horses treated with ivermectin or pyrantel pamoate, and during the second day from horses treated with fenbendazole. Fifteen cyathostomin species were identified and the six most prevalent were Cylicocyclus nassatus, Cyathostomum catinatum, Cylicostephanus longibursatus, Cylicocyclus leptostomus, Cylicostephanus minutus and Cylicostephanus calicatus. These species composed 91% of the total burden of cyathostomins. The number of species found per horse ranged from 6 to 13, with an average of 9. No significant differences in species composition or distribution were found between the treatment groups. On day 5, i.e. 1 day after the last ivermectin treatment, 93% of the adult worms were recovered from horses in the fenbendazole group.This study showed that it was possible to identify cyathostomins expelled in faeces of dewormed horses, and that the most prevalent species corresponded to those found in autopsy surveys performed in other countries.     

Farming practices in Sweden related to feeding milk and colostrum from cows treated with antimicrobials to dairy calves

Background

Milk produced by cows in receipt of antimicrobial therapy may contain antimicrobial residues. Such antimicrobial-containing waste milk must be withdrawn from human consumption and is therefore sometimes used as calf feed. Unfortunately, this approach might promote selection of antimicrobial resistant bacteria in the calves’ intestinal microbiota. The objectives of this study were therefore to obtain an overview of waste milk feeding practices on Swedish dairy farms and to investigate if these practices were associated with certain farm characteristics. A representative group of 457 Swedish dairy farmers participated in a web-based survey with questions about the use of colostrum and milk from cows treated with antimicrobials at dry off or during lactation, respectively, as calf feed.

Results

Colostrum (milk from the first milking after calving) and transition milk (milk from the second milking to the fourth day after calving) from cows treated with antimicrobials at dry off was fed to calves on 89% and 85% of the farms in the study, respectively. When antimicrobial therapy was given to cows during lactation, 56% of the farms fed milk that was produced during the course of treatment to calves, whereas milk that was produced during the subsequent withdrawal period was fed to calves on 79% of the farms. Surveyed farmers were less prone to feed such milk if the antimicrobial therapy was due to mastitis than other infections. In Sweden, a majority of antimicrobial treatments during lactation are systemic administration of benzylpenicillin and thus, the bulk of waste milk in Sweden is likely to contain residues of this drug. Feeding waste milk to calves was more common on non-organic farms, and on farms located in Southern Sweden, and was less common on farms with cows housed in cold free stalls barns.

Conclusions

Waste milk that may contain antimicrobial residues is, at least occasionally, used as feed for calves on a majority of surveyed Swedish dairy farms. Future work should focus on the effect of waste milk feeding on the occurrence of antimicrobial resistant bacteria in the calves’ intestinal microbiota.     

Estimating the biomass and carbon pool of stump systems at a national scale

Countries that are signatories to the UNFCCC and its supplementary Kyoto Protocol are obliged to report changes in carbon pools. These should include the pool of carbon held in tree stumps and roots but, to date, few countries have been able to report this or separate it from the dead-wood pool. The aim of this study was to develop a general system for estimating and monitoring changes in stump system carbon using data from a traditional National Forest Inventory. The system was derived using data based on measurements of carbon (biomass) in inventoried permanent sample plots representing all relevant classes of land-use. With this design it was possible to trace matched carbon at the level of individual trees or stumps back to land-use prior to the 1990 baseline year. Between 1990 and 2003 in Sweden, the average annual net sink of stump systems was estimated to amount to 6.7 Mt CO₂ equiv. year⁻¹ – comparable to the reported net sink in 2008 of about 15 Mt CO₂ equiv. year⁻¹ from the whole Land Use, Land-Use Change and Forestry sector, which excluded any carbon in stump systems. In 2003 the carbon stock of stumps and roots was estimated at 495 Mt CO₂ equiv.; approximately five times that of the dead-wood pool as defined in Sweden, i.e. dead wood that mainly consists of boles. The Intergovernmental Panel on Climate Change requests that reported carbon should be matched to land-use and traced back to the 1990 base year; however, the present study confirms expectations that most carbon in stumps and roots is found on Forest land. The minimum requirements for estimating the carbon pool in stump systems at a national scale using the proposed methodology are that there should be: (i) a consistent time-series of harvest data, usually estimated as merchantable volume; (ii) conversion factors from merchantable volume to stump system biomass at death; and (iii) a representative decomposition model.     

Encephalomyelitis and myositis in a boxer puppy due to a Neospora-like infection

A 10-week-old Boxer dog from Uppsala, Sweden, was killed because of hind limb paralysis. The hind limb muscles were grossly atrophied and encephalomyelitis and myositis were the predominant microscopic lesions. Neospora caninum-like tachyzoites were found in the brain and skeletal muscles, and tissue cysts were found in the brain. The organism was not Toxoplasma gondii, as T. gondii antibody was not found in the indirect fluorescent antibody test in serum samples from the puppy and the dam, and the organism in the dog's brain did not react with T. gondii antibody in the immunoperoxidase test.     

A long-term study of Neospora caninum infection in a Swedish dairy herd

A longitudinal study was performed in a Swedish dairy herd where Neospora caninum had been isolated from a stillborn calf. Starting in autumn 1994, blood samples from all female animals in the herd were collected once yearly until 1999. The sera were analysed for presence of IgG1 antibodies to N. caninum by the iscom ELISA, and by an avidity ELISA to establish the timing of infection. In addition, data on reproductive performance were compiled. During the study the percentage of seropositive female animals increased from 63% to 87%. In 1994 a large number of young animals tested seropositive although their dams were seronegative, indicating that a transmission of the parasite other than the vertical had recently occurred. Low avidity values supported this assumption. The annual abortion rate increased from a mean of 2% before the initiation of the study to 9% in 1994-1998. During the same time, as judged by the avidity data, a large proportion of the animals shifted from being recently to being chronically infected. The source of the external infection in the herd could not be identified.     

A 3-year field evaluation of pasture rotation and supplementary feeding to control parasite infection in first-season grazing cattle: dynamics of pasture infectivity

A 3-year grazing trial (2002-2004) was conducted on a commercial beef cattle farm in south-central Sweden to assess different methods of parasite control. This paper focuses on the dynamics of the free-living larval stages, whereas data on performance and within-host parasitological variables are presented in a complementary paper. Each year in May, 4 groups of 10 first-season grazing (FSG) steers were turned out on to separate 2ha paddocks and subjected to the following strategies: (1) spring turn-out on to pasture which had been grazed the previous year by second-season grazing (SSG) steers (paddock RT), followed by a move to aftermath (paddock AM) after 10 weeks (mid-July), (2) supplementary feeding with concentrate and hay for 4 weeks following turn-out (paddock FD), set stocked, (3) untreated control (paddock UT), set stocked and (4) anthelmintic treated control (paddock DO), set stocked. All paddocks were assigned a new set of FSG cattle each year whereas the treatments remained the same. Pasture infectivity were monitored partly by two tracer calves that grazed each paddock along with the FSG calves for 3 weeks after turn-out and prior to housing, partly by analysis of herbage samples for infective larvae (L3) that were collected from each paddock at monthly intervals between April and October. The predominant genera found were Cooperia and Ostertagia. Tracers grazing paddock RT overall harboured less worms, and in particular less Ostertagia spp., and tracers grazing paddock AM in mid-July harboured insignificant numbers of nematodes compared to tracers on the FD and UT paddocks. Although total worm counts varied between groups, smaller numbers were generally observed early in the grazing-season (May), compared to close to housing (September) when inhibited early L4 larvae were almost exclusively found. Results observed from herbage samples showed high numbers of L3 in spring before the time of turn-out, compared to around housing. In conclusion, the rotation control strategy showed promising results and provided a turn-out pasture that was 'nematode safe' to FSG cattle the following spring, whereas the feeding strategy failed as applied in this experiment.     

A Field Study on the Effect of Some Anthelmintics on Cyathostomins of Horses in Sweden

The objective of the study was to investigate different aspects on the efficacy of three anthelmintics on cyathostomin nematodes of Swedish horses. A faecal egg count reduction (FECR) test was performed on 26 farms. Horses were treated orally with recommended doses of ivermectin, pyrantel pamoate or fenbendazole. Faecal samples were collected on the day of deworming and 7, 14 and 21 days later. No resistance was shown against ivermectin; the FECR was constantly >99%. The effect of pyrantel was assessed as equivocal in 6 farms 14 days after treatment; the mean FECR was 99%. As many as 72% of the fenbendazole-treated groups met the criteria for resistance; the mean FECR was 86%, ranging from 56% to 100%. A re-investigation of two farms where pyrantel resistance had been suspected clearly revealed unsatisfactory efficacy of pyrantel on one of these farms; the FECR varied from 72% to 89%. Twenty-six of the horses previously dosed with pyrantel or fenbendazole, and which still excreted ≥150 eggs per gram of faeces 14 days after treatment, were dewormed with ivermectin and fenbendazole or pyrantel in order to eliminate the remaining cyathostomins. A total of 13 cyathostomin species were identified from horses that initially received fenbendazole and seven species were identified from pyrantel-treated individuals. The egg reappearance period (ERP) following treatment with ivermectin and pyrantel was investigated on two farms. The shortest ERP after ivermectin treatment was 8 weeks and after pyrantel was 5 weeks. We conclude that no substantial reversion to benzimidazole susceptibility had taken place, although these drugs have scarcely been used (<5%) in horses for the last 10 years. Pyrantel-resistant populations of cyathostomins are present on Swedish horse farms, but the overall efficacy of pyrantel is still acceptable.     

Infectivity and cross-immunity studies of Theileria lestoquardi and Theileria annulata in sheep and cattle: I. In vivo responses.

In a series of experiments, sporozoite stabilates of a Theileria lestoquardi (Lahr) and a T. annulata (Ankara) stock prepared from Hyalomma anatolicum anatolicum ticks, were used to examine the infectivity of both parasite species for sheep and cattle and to study the development of cross-immunity between these parasite species. In the first experiment sheep and cattle were inoculated with T. lestoquardi sporozoites. Surviving animals and naive sheep and cattle were, in the second experiment, inoculated with T. annulata. In the third experiment, naive sheep and sheep previously infected with T. annulata, were inoculated with T. lestoquardi. The following responses to inoculations were monitored: clinical and haematological signs of infection, appearance of parasitic stages of the parasites in lymph node biopsies and in peripheral blood and serological response to T. lestoquardi and T. annulata schizont antigens. While T. lestoquardi readily infected sheep and caused severe disease, it did not infect cattle. On the other hand, T. annulata infected both cattle and sheep. However, whereas cattle became severely affected, infected sheep showed mild clinical symptoms only and piroplasms did not develop. Despite their different behaviour in the host species examined, cross-immunity studies suggested that the parasite species are very closely related. Experiments in sheep indicated that T. lestoquardi infection protected against subsequent T. annulata infection. On the other hand, recovery from T. annulata infection did not prevent infection by sporozoites of T. lestoquardi, resulting in the establishment of schizonts and their subsequent development into piroplasms, although it protected against the major clinical effects of T. lestoquardi infection.