An assay for quantitative virulence in Rhynchosporium commune reveals an association between effector genotype and virulence

Detailed knowledge of the evolutionary genetics of virulence is needed to understand and predict host–pathogen dynamics. This study used a virulence assay based on digital image analysis and treated virulence as a quantitative rather than a binary trait. Such quantitative data may better reflect the genetic underpinning of virulence in many pathogen systems and provide better resolution in statistical investigations. A greenhouse experiment based on a common garden design was conducted to measure virulence (% of leaf area covered by lesions) of 126 genetically distinct isolates of the barley scald pathogen, Rhynchosporium commune, originating from nine field populations around the world. Virulence in this pathosystem was found to be a quantitative trait with a continuous distribution in all populations. By comparing population genetic differentiation for virulence and neutral microsatellite markers (i.e. a Q_ST/G_ST comparison), evidence that virulence is under stabilizing selection across populations was found. Heritability values were high and ranged from 0·52 to 0·96 with a mean heritability of 0·84. Virulence was positively correlated with spore production as predicted by the trade‐off theory of virulence evolution. Furthermore, an association analysis between virulence and sequence haplotypes of three known necrosis‐inducing effector genes (NIP1, NIP2 and NIP3) revealed a significant effect of NIP2 haplotypes and NIP1 deletions. Overall, the results support a quantitative model for virulence in the R. commune–barley pathosystem and very high evolutionary potential for this trait.     

CNS mastitis: nothing to worry about?

In this paper, we analyzed a very large field data set on intramammary infections (IMI) and the associated somatic cell count (SCC) in dairy cows. The objective of the study was to analyze the impact of coagulase-negative staphylococci (CNS) IMI on cow SCC, both mean and variability, and on the potential of these infections to have a major impact on the bulk milk SCC (BMSCC). Data and milk samples for bacterial culture were collected by Quality Milk Production Services (QMPS) between 1992 and March of 2007. The QMPS program services dairy farms in New York State and other states in the Northeastern USA and operates in conjunction with Cornell University. Only records from cows where SCC and milk production data were available, and where only one organism was isolated from bacterial cultures of milk samples (or where culture was negative) were used for this analysis. A total of 352,614 records from 4200 whole herd mastitis screening sampling qualified for this study. Within herds an average of 15% (S.D. 12%) of cows sampled were infected with CNS, ranging between 0 and 100%. Average within herd prevalence of cows with a CNS IMI and an SCC over 200,000 cells/ml was 2% (S.D. 4%) with a minimum of 0% and a maximum of 50%. Results of linear mixed models showed three distinct populations of IMI statuses: negative cultures with the lowest SCC; CNS and Corynebacterium bovis with a moderate increase in SCC, and Streptococcus agalactiae, Streptococcus spp. and Staphylococcus aureus showing an important increase in SCC. Surprisingly, milk production was slightly but significantly higher in CNS infected cows compared to culture-negative cows, whereas it was strongly reduced in cows with a major pathogen IMI. The percentage contribution of CNS infections to the BMSCC was 17.9% in herds with a BMSCC less than 200,000 cells/ml. This value decreased to 11.9 and 7.9% in herds with bulk milk SCC between 200,000 and 400,000 and over 400,000 cells/ml, respectively. We concluded that very few herds with milk quality problems would have an important increase in BMSCC that could be mostly attributed to CNS infections. On the other hand, in herds with low BMSCC, CNS infections may be an important contributor to the total number of somatic cells in the bulk milk.     

Evaluation of hemostatic defects secondary to vascular tumors in dogs: 11 cases (1983-1988)

Two populations of dogs with cutaneous hemangiomas and hemangiosarcomas were evaluated retrospectively. One population consisted of 96 dogs seen at the Veterinary Medical Teaching Hospital at the University of California, Davis. The second population consisted of 116 dogs that had skin biopsy specimens submitted to a private veterinary diagnostic laboratory for histologic diagnosis. Nine dogs from the teaching hospital and 2 dogs, from which samples had been submitted to the veterinary diagnostic laboratory, developed hemostatic defects in association with the tumors. Hemostatic defects included hemorrhage directly from the tumor, thrombocytopenia, hypofibrinogenemia, and findings associated with disseminated intravascular coagulation. Because bleeding during surgery can develop in animals with hemostatic defects, dogs with one or more tumors suspected of being vascular in origin should have platelet numbers and hemostatic analytes evaluated prior to surgery, especially if petechiae or ecchymoses are evident.     

The effect of feeding level and physiological status on total flow and amino acid composition of endogenous protein at the distal ileum in swine.

An experiment was conducted to investigate the effects of BW, feed intake, and the physiological condition of the animal on the loss and amino acid composition of endogenous protein in swine. Ten growing barrows and five multiparous sows were equipped with a T-cannula in the distal ileum for digesta collection. A protein-free diet was fed to all animals. The barrows were given free access to the experimental diet. The sows also were allowed to consume the diet on an ad libitum basis, and digesta were collected during lactation and in the following gestation period. In addition, digesta from the gravid sows were collected after restricting the sows to 2 kg of feed per day. For each animal group, the endogenous losses of protein and amino acids were calculated in relation to DMI, and the amino acid composition of endogenous protein was calculated. The total endogenous gut protein loss at the distal ileum of growing pigs, lactating sows, and gestating sows, given free access to feed, was 12.4, 9.4, and 11.2 g/kg DMI, respectively. These values were not different (P > .10). However, when gestating sows were fed only 2 kg/d, 17.8 g of endogenous protein was lost per kilogram of DMI, which was higher (P < .05) than for any of the other groups. This difference was mainly caused by higher (P < .05) losses of glycine, proline, and serine. There were no differences (P > .05) in amino acid composition of endogenous protein between growing pigs, lactating sows, and gestating sows given free access to feed, but restricted-fed gestating sows had an amino acid composition of endogenous protein that was significantly different from that of the other groups. The results from the experiment showed that age, BW, and the physiological condition of the animal have little or no effect on the amount of endogenous protein and amino acids lost at the distal ileum of hogs if calculated in relation to DMI. Likewise, the amino acid composition was not affected by the BW or physiological condition of the animal. However, DMI had a significant effect on endogenous protein losses in sows as well as on amino acid composition of endogenous protein.     

Water balance in desert arthropods. Despite their small size, arthropods may be highly adapted for life in xeric conditions

As judged by the number of species, or of individuals, arthropods are an extremely successful group of desert inhabitants. There is very great structural and physiological diversity within the group, and since adaptations to desert life open to one are not open to all. we should not expect to find the maximum possible development of adaptive features in any arthropod simply because it lives in a desert. Most adult insects fly; their larvae and all other arthropods do not, and their adaptations will differ accordingly. Desert beetles have very impermeable cuticles and tolerate high body temperatures, while desert cockroaches live below the sand. have more permeable cuticles, and absorb water vapor.There is probably no single respect in which all desert arthropods differ from insects of other environments. Perhaps a profitable way of viewing desert animals is to recognize that each is a whole organism with a specific collection of adaptations that must be consistent within themselves and which are associated with a specific mode of life and a specific evolutionary history. The arthropod organization is capable of producing highly efficient desert species. There is, however, a converse way of looking at the situation, Which is often neglected but which may be of general biological interest: does the evolution of adaptations to desert environments necessarily involve loss of viability in more mesic habitats? If so, then what are these disavantages- what, for example, is the disadvantage of a highly impermeable cuticle? In some cases the answer is clear: sandroaches need sand dunes to live in because they are morphologically and behaviorly specialized for this habitat. More often the answer is not obvious.     

Qualitative tear film and conjunctival goblet cell assessment of cats with corneal sequestra

OBJECTIVES: To evaluate the tear film qualitatively and conjunctival goblet cell numbers in cats with and without corneal sequestra. ANIMALS STUDIED AND PROCEDURES: This was a prospective evaluation of 11 cats with corneal sequestra and 14 control eyes that were either the contralateral normal eye when the sequestrum was unilateral or from control cats of similar age with no ocular disease. All cats in this study were examined by a veterinary ophthalmologist. The ophthalmic examinations included a neuro-ophthalmic evaluation, Schirmer tear tests, fluorescein staining, tear film break-up times, applanation tonometry, biomicroscopy, and indirect ophthalmoscopy. The palpebral conjunctiva at the dorsal nasal, ventral nasal, dorsal temporal and ventral temporal fornices were biopsied after topical anesthetic was applied to the cornea and conjunctiva. The conjunctival biopsies were fixed in formalin and sectioned routinely and stained with hematoxylin and eosin, and periodic acid-Schiff. These slides were examined by light microscopy by a blinded examiner. Goblet cell numbers were compared to conjunctival basal epithelial cell numbers by region. The goblet cell numbers by region from the eyes with sequestra was statistically compared to those from eyes without sequestra, with a student's paired t-test. Conjunctival swabs were collected from the cats with corneal sequestra and submitted for polymerase chain reaction for Herpes felis, Chlamydia psiitticia, and Mycoplasma felis. The corneal sequestra were removed by surgical keratectomy and fixed and stained routinely, and examined by light microscopy. RESULTS: No neurologic abnormalities were detected in any of the cats. The Schirmer tear tests (eyes with sequestra 14+/-5.1 mm/min; normal eyes 15+/-6.8 mm/min) and intraocular pressures (eyes with sequestra 21+/-6.6; normal eyes 22+/-5.8) were within normal reference ranges for cats. Biomicroscopic examinations revealed varied sizes and depths of brown- and amber-colored corneal sequestra. No abnormalities were noted on indirect ophthalmoscopic examinations. The tear film break-up time was 21 s (+/-12) for the normal eyes (n=14) and 14 s (+/-13) in eyes with corneal sequestra (n=11). The average goblet/epithelial cell ratios by region for the normal eyes and the eyes with sequestra respectively were 0.66, 0.56 for the dorsal nasal fornix, 0.68, 0.57 for the ventral nasal fornix, 0.63, 0.48 for the temporal dorsal fornix, and 0.55, 0.49 for the temporal ventral fornix. There were no significant differences in tear film break-up times and goblet cell numbers in eyes with corneal sequestra and those without sequestra. Three conjunctival swabs from two of 11 cats with sequestra were positive with PCR for Herpes felis virus. These included one cat with bilateral sequestra and one cat with unilateral corneal sequestrum. CONCLUSIONS: The pathogenesis of feline corneal sequestra does not appear to be linked primarily to abnormal goblet cell numbers, qualitative tear film abnormalities, and accelerated tear film break-up time.     

Spatial Clustering of Escherichia coli with Reduced Susceptibility to Cefotaxime and Ciprofloxacin among Dairy Cattle Farms Relative to European Starling Night Roosts

European starlings (Sturnus vulgaris) have been implicated in the dispersal of zoonotic enteric pathogens. However, their role in disseminating antimicrobial‐resistant organisms through their home range has not been clearly established. The aim of this study was to determine whether starling night roosts served as foci for spreading organisms with reduced susceptibility to antimicrobials among dairy cattle farms. Bovine faecal pats were collected from 150 dairy farms in Ohio. Each farm was visited twice (in summer and fall) between 2007 and 2009. A total of 1490 samples (10 samples/farm over two visits) were tested for Escherichia coli with reduced susceptibility to cefotaxime and ciprofloxacin. Using a spatial scan statistic, focal scans were conducted to determine whether clusters of farms with a high prevalence of organisms with reduced susceptibility to cefotaxime and ciprofloxacin surrounded starling night roosts. Faecal pats 13.42% and 13.56% of samples carried Escherichia coli with reduced susceptibility to cefotaxime and ciprofloxacin, respectively. Statistically significant (P < 0.05) spatial clusters of faecal pats with high prevalence of Escherichia coli showing reduced susceptibility to cefotaxime and ciprofloxacin were identified around these night roosts. This finding suggests that the risk of carriage of organisms with reduced susceptibility to antimicrobials in cattle closer to starling night roosts was higher compared to cattle located on farms further from these sites. Starlings might have an important role in spreading antimicrobial‐resistant E. coli to livestock environments, thus posing a threat to animal and public health.     

Diagnosis of ovine fascioliasis by a dot enzyme-linked immunosorbent assay: a rapid microdiagnostic technique

A microenzyme-linked immunosorbent assay (dot-ELISA) was modified for making an immunodiagnosis of Fasciola hepatica infections in sheep. Sheep were alloted as follows: group I-3 controls and 4 principals, each inoculated with 500 metacercariae; group II-3 controls and 7 principals, each inoculated with 250 metacercariae; and group III-3 controls and 7 principals, each inoculated with 500 metacercariae. Blood and fecal samples were collected from each animal every 2 weeks for 16 weeks. Presence (or absence) of flukes was confirmed by fecal examinations and examination of dissected livers at necropsy of the sheep. The dot-ELISA incubations were done at ambient room temperature. Nitrocellulose disks dotted with 1 microliter (50 ng of protein) of F hepatica excretory/secretory products were placed in 96-well tissue culture plates. After nonspecific binding sites on the disks were bound with bovine serum albumin-triethanolamine-buffered saline solution, dilutions (1:2) of positive- and negative-control serum samples or experimental serum samples were placed in appropriate wells for a 30-minute incubation. Wells were washed (3 times), and 50 microliters of horseradish peroxidase conjugated rabbit anti-sheep immunoglobulin G was added to each well for a 30-minute incubation and then aspirated. Substrate solution (4-chloro-1-naphthol, methanol, triethanolamine-buffered saline solution, and H2O2; 50 microliters) was added for a 30-minute incubation and then aspirated. Disks were air dried for visualization: solid purple dot = positive sample, or no dot = negative sample.(ABSTRACT TRUNCATED AT 250 WORDS)