Inter-rater agreement in visual assessment of footpad dermatitis in Danish broiler chickens.

1. The performance of the scoring in the Danish footpad dermatitis (FPD) surveillance system was evaluated by determining inter-rater agreement in visual inspection of FPD in broilers between two independent raters (R1 and R2) and the official scoring at a Danish slaughterhouse. 2. FPD scores were evaluated in 1599 chicken feet. The two raters and the slaughterhouse scored equal proportions of score 0. So did R1 and R2 when assessing score 1 and the more severe lesion score 2, whereas the slaughterhouse scored a markedly higher proportion of score 1 and a lower proportion of score 2. Aggregated FPD flock scores ranged from 5 to 163 (R1 and R2) and from 8 to 107 (slaughterhouse). 3. The level of agreement between the two raters was high for scores 0, 1 and 2 and for flock scores. Agreement between raters and the slaughterhouse was lower when R1 and R2 recorded score 2 than when they recorded scores 0 and 1. 4. This study indicates that the occurrence and severity of lesions are underestimated in the official Danish FPD scoring system.

Evaluation of the Danish footpad lesion surveillance in conventional and organic broilers: Misclassification of scoring.

Danish legislation prescribes surveillance of footpad dermatitis (FPD) at slaughter as an indicator of on-farm broiler welfare. The 3-point scale being used was originally developed in Sweden to score feet from conventional broilers, but the extent and causes of misclassifications have not been investigated, neither in conventional nor organic broilers. Hence, we investigated the performance of the official Danish FPD scoring system in conventional and organic broilers by assessing agreement between official scores from the slaughterhouse and consecutive scoring of the same feet by a reference method. We also investigated the impact of performing an incision of the footpad during scoring. In total, 902 conventional and 897 organic broiler feet (∼100 per flock from 18 flocks) were collected at a large Danish slaughterhouse for the official FPD surveillance system. Laboratory scoring, according to predefined criteria for visual and invasive investigations of the feet derived from the official system, was compared to the official scores assigned at slaughter. Footpad lesions were primarily chronic, representing a wide range of severity. Marked differences in color, shape, and degree of papillary hypertrophy and hyperkeratosis of organic and conventional feet were observed. Low agreement primarily regarding score 2 lesions was observed when comparing official and reference foot scores in conventional (0.31) and organic (0.05) broilers. Variation in agreement when comparing flock scores suggested a non-systematic bias, which might be attributed to differences among official raters. The very low agreement in feet from organic broilers shows that these were more difficult to score than conventional. This might be due to a mismatch between lesion characteristics and scoring criteria, or because the lesions were less severe. Strictly visual examination detected 3 out of 4 score 2 lesions identified by the reference method. This study indicated that a large proportion of severe FPD lesions go unnoticed in the official Danish scoring system. The results further suggested that the complexity and impracticality of the scoring criteria impede uniform scoring among raters and production systems.

Sorbitol non-fermenting shiga toxin-producing Escherichia coli in cattle on smallholdings.

We investigated faecal samples collected from the rectum of 518 cattle on 371 randomly selected smallholdings in Bangladesh for the presence of sorbitol non-fermenting (SN-F) shiga toxin-producing Escherichia coli (STEC). The SN-F isolates were tested for the presence of rfb O157, stx1, stx2, eae and hlyA genes by polymerase chain reaction (PCR). Seven SN-F isolates lacking these genes were profiled by pulsed-field gel electrophoresis (PFGE) to verify their clonality. SN-F E. coli was identified in 44 [8·5%, 95% confidence interval (CI) 6·4-11·2] samples; of these, 28 (5·4%, 95% CI 3·8-7·7) had shiga toxin-producing strains, although only two carried the rfb O157 gene. Thirteen isolates carried the hlyA gene while 18 harboured the eae gene. Based on PFGE, six pulsotypes were observed among the seven isolates that had no virulence genes. To the best of our knowledge this is the first report on shiga toxin-producing E. coli from direct rectal faecal samples of cattle on smallholdings.

IFNγ and perforin cooperate to control infection and prevent fatal pathology during persistent gammaherpesvirus infection in mice.

Infection with murine gammaherpesvirus 68 has become an accepted model for studying the virus/host interactions with regard to gammaherpesvirus infections. Previous studies using gene-deficient mice have revealed that neither IFNγ nor perforin is essential in controlling the outcome of infection or the virus load during chronic infection in C57BL/6 mice. However, pronounced multiorgan fibrosis and splenic atrophy are observed in mice lacking IFNγ or the IFNγ receptor. To study the interplay between perforin and IFNγ in controlling the virus-induced pathology and the viral load during chronic gammaherpesvirus infection, we infected IFNγ/perforin double-deficient C57BL/6 mice and followed the course of infection. While absence of perforin prevented the splenic atrophy in IFNγ-deficient mice, fibrosis did not disappear. Moreover, double-deficient mice developed extreme splenomegaly, were unable to control the viral load and displayed chronic immune activation. Thus, IFNγ and perforin act in concert to minimize pathology and control the viral load in mice chronically infected with MHV68. Furthermore, while certain aspect of the virus-induced pathology in IFNγ-deficient mice may be alleviated in double-deficient mice, other aspects are exaggerated, and the normal architecture of the spleen is completely destroyed. We believe that these findings add to the understanding of the virus/host interaction during chronic gammaherpes virus infection.

Pathological manifestations observed in dead-on-arrival broilers at a Danish abattoir.

1. The mortality of broilers during pre-slaughter handling, including harvesting and transport, is an issue of increasing public concern which has led to the adoption of Council Directive EC/43/2007 implementing abattoir surveillance regarding the number of dead-on-arrival (DOA) broilers. 2. Pathological lesions and causes of death of DOA broilers at a Danish abattoir were investigated in a cross-sectional study comprising 300 DOA broilers (25 broilers from each of 12 randomly selected flocks). Major pathological manifestations of DOA broilers included severe pulmonary congestion (51.5%), lung congestion in combination with trauma (12.5%), trauma (10.2%), nephropathy accompanied by dehydration and/or discolouration (8.8%), morbus cordis (2.0%), septicaemia (1.7%) and suspected septicaemia (1.0%). Lung congestion accompanied by circulatory disturbances in other tissues was suggested to be due to suffocation. 3. Analyses of pathological diagnoses revealed that DOA broilers can be divided into two main categories, lung congestion and trauma, based on the chronicity of the lesions, both of which are primarily related to management and handling procedures. Most DOA broilers examined (74.2%) were estimated to have died as a consequence of events during pre-slaughter handling underlining the importance of increased focus on handling-related factors to reduce DOA rate.

Antihistomonal effects of artemisinin and Artemisia annua extracts in vitro could not be confirmed by in vivo experiments in turkeys and chickens.

Five different Artemisia annua-derived materials (i.e. dry leaves, pure artemisinin, and hexane, dichloromethane or methanol extracts of leaves) were screened for their in vitro activities against six clonal cultures of Histomonas meleagridis. Except for the methanol extract, all tested materials displayed in vitro activity against all tested protozoal clones. Neither the dry plant material, extracts nor artemisinin showed any antibacterial activity against the xenic bacteria accompanying the six H. meleagridis clones at concentration levels identical to the antihistomonal setting. The dichloromethane extract of dry leaves (Ext-DCM) (minimal lethal concentration=1.0 mg/ml) and artemisinin (half-maximal inhibitory concentration=1.295 mg/ml) had the most promising antihistomonal properties and were therefore subsequently tested in a standardized experimental infection model in both turkeys and chickens infected with clonal H. meleagridis. There were no differences between treatment groups, where all infected turkeys showed severe clinical histomonosis and demonstrated severe typhlohepatitis typical for histomonosis. Consistent with the infection model used, the infected chickens did not show any adverse clinical signs but contracted severe lesions in their caeca 7 and 10 days post infection (d.p.i.), liver lesions were absent to mild after 7 d.p.i. and progressed to severe lesions at 10 d.p.i.; thus no differences between treatment groups were observed. In conclusion, neither artemisinin nor Ext-DCM was able to prevent experimental histomonosis in turkeys and chickens at the given concentrations, which is contrary to the antihistomonal effect noticed in vitro even though the same clonal culture was used. The results of this study therefore clearly demonstrate the importance of defined in vivo experimentation in order to assess and verify in vitro results.

Risk factors associated with anthrax in cattle on smallholdings.

SUMMARY Unprecedented high rates of anthrax outbreaks have been observed recently in cattle and humans in Bangladesh, with 607 human cases in 2010. By enrolling 15 case and 15 control cattle smallholdings in the spatial zone in July-September 2010, we conducted a case-control study, data of which were analysed by matched-pair analysis and multivariable conditional logistic regression. Feeding animals with uprooted and unwashed grass [odds ratio (OR) 41·2, 95% confidence interval (CI) 3·7-458·8, P=0·003], and feeding water hyacinth (Eichhornia crassipes) (OR 22·2, 95% CI 1·2-418·7, P=0·039) were independent risk factors for anthrax in cattle.

Clonality and virulence traits of Escherichia coli associated with haemorrhagic septicaemia in turkeys.

Fifty-five clinical isolates of avian pathogenic Escherichia coli (APEC) from seven outbreaks of acute haemorrhagic septicaemia in turkeys were characterized by serotyping, plasmid profiling including restriction analysis with HindIII, ribotyping with EcoRI and HindIII, multilocus sequence typing (MLST) and virulence profiling. A clonal relationship was demonstrated for each outbreak according to serotype, plasmid profiling, ribotyping, and MLST. In addition, isolates demonstrated highly similar virulence profiles, as all isolates were positive for F11 pili and possessed genes encoding aerobactin (iucD), increased serum survival (iss), temperature-sensitive haemagglutinin (tsh) and colicin V plasmid operon genes (cva/cvi). However, only 20% of the isolates produced colicin V and 42% exhibited serum resistance. All strains with O group O111 and a single O18ac strain (demonstrating non-clonal DNA profiles) were positive for enteroaggregative heat-stabile toxin (EAST1), while isolates of a single outbreak all possessed the enteroaggregative toxin gene (astA). All isolates were negative for genes encoding verocytotoxins (vtx/stx), iron-repressible protein (irp2), P-fimbria (papC), invasion plasmid antigen (ipaH), attaching and effacing gene (eae), enterohaemolysin (ehxA), and enterotoxins LT, STIa (ST(p)) and STIb (ST(h)). In conclusion, highly similar virulence profiles were demonstrated for isolates of E. coli associated with a single well-defined lesion type of colibacillosis in turkeys; acute haemorrhagic septicaemia. The isolates obtained, however, demonstrated a different phylogenetic background, underlining the importance of using well-defined strain collections for characterization of APEC pathotypes.

Time-course investigation of infection with a low virulent Pasteurella multocida strain in normal and immune-suppressed 12-week-old free-range chickens.

Twelve-week-old indigenous chickens, either immune-suppressed using dexamethasone (IS) or non-immune-suppressed (NIS), were challenged with a low virulent strain, Pasteurella multocida strain NCTC 10322(T), and developed clinical signs and pathological lesions typical of chronic fowl cholera. NIS birds demonstrated much more severe signs of fowl cholera than IS birds. With few exceptions, signs recorded in IS and NIS birds were of the same types, but significantly milder in the IS birds, indicating that immune suppression does not change the course of infection but rather the severity of signs in fowl cholera. P. multocida signals by fluorescent in situ hybridization (FISH) were observed between 1 h and 14 days in the lungs, trachea, air sacs, liver, spleen, bursa of Fabricius and caecal tonsils, while signals from other organs mostly were observed after 24 h. More organs had FISH signals in NIS birds than in IS birds and at higher frequency per organ. Many organs were positive by FISH even 14 days post infection, and it is suggested that these organs may be likely places for long-term carriage of P. multocida following infection. The present study has demonstrated the spread of P. multocida in different tissues in chickens and distribution of lesions associated with chronic fowl cholera, and pointed to a decrease of pathology in IS birds. Since dexamethasone mostly affects heterophils, the study suggests that these cells play a role in the development of lesions associated with chronic fowl cholera in chickens.

Multi-locus sequence typing and plasmid profile characterization of avian pathogenic Escherichia coli associated with increased mortality in free-range layer flocks.

Avian pathogenic Escherichia coli strains originating from 10 free-range layer flocks were characterized by multi-locus sequence typing and plasmid profile analysis to investigate their phylogenetic relationship and diversity, respectively. In addition to colibacillosis, all flocks tested positive for antibodies against avian metapneumovirus (aMPV) during production, and six of the flocks were concurrently affected by histomonosis. Accumulated average mortality for flocks concurrently affected by colibacillosis and histomonosis made up 17.4%, while the average mortality for E. coli-infected flocks was 16.5%. A total of eight different sequence types (STs) and 47 different plasmid profiles were demonstrated among the E. coli isolates. Within each flock between one and four different STs and between three and 13 different plasmid profiles were demonstrated. A statistical significant difference in STs and plasmid profile diversity of the population of E. coli was not demonstrated between flocks affected by histomonosis compared with histomonosis-free flocks. Only minor clonal diversity was demonstrated for each flock, and in all but one flock colibacillosis started before antibodies against aMPV were detected. All isolates, except two, carried plasmids greater than 100 kb, but only a single plasmid replicon type, IncFIB, was demonstrated, suggesting plasmids representing this type might represent a common pathogenicity factor for the different STs of E. coli. Within each flock a clonal tendency was observed, indicating that only certain clones of E. coli possess a significant pathogenic potential. These clones act as primary rather than secondary pathogens, resulting in colibacillosis without predisposing factors, including histomonosis and aMPV.

Risk for highly pathogenic avian influenza H5N1 virus infection in chickens in small-scale commercial farms, in a high-risk area, Bangladesh, 2008.

Small-scale commercial chicken farms (FAO-defined system 3) with poor biosecurity predominate in developing countries including Bangladesh. By enroling fifteen highly pathogenic avian influenza (HPAI) cases occurring in such farms - (February - April 2008) and 45 control farms (March-May 2008) with similar set up, we conducted a case-control study to evaluate the risk factors associated with HPAI H5N1 virus infections in chickens reared in small-scale commercial farms in a spatially high-risk area in Bangladesh. Data collected by a questionnaire from the selected farms were analysed by univariable analysis and multivariable conditional logistic regression. The risk factors independently associated were 'dead crow seen at or near farm' [odds ratio (OR) 47.4, 95% confidence interval (CI) 4.7-480.3, P = 0.001], 'exchanging eggtrays with market vendors' (OR 20.4, 95% CI 1.9-225.5, P = 0.014) and 'mortality seen in backyard chicken reared nearby' (OR 19.4, 95% CI 2.8-131.9, P = 0.002). These observations suggest that improved biosecurity might reduce the occurrence of HPAI outbreaks in small-scale commercial farms in Bangladesh.

Mortality rate and clinical features of highly pathogenic avian influenza in naturally infected chickens in Bangladesh.

A retrospective study was performed to assess the mortality rates and clinical signs in commercial and backyard chickens naturally infected with the highly pathogenic avian influenza (HPAI) H5N1 virus in Bangladesh. All of the 33 commercial and 25 backyard farms, recorded during the first wave of HPAI outbreaks (January to November 2007) in Bangladesh, were enrolled in this study. The farm profile data, stock information and major clinical signs recognised by the farmers and/or farm attendants, were collected through a prototype questionnaire. The investigating veterinarians' retrospective appraisals of the clinical signs were recorded by interview. After the clinical onset, the incidence rates of mortality in breeder, commercial broiler, commercial layer and backyard chickens were, respectively: 0.0215, 0.0341, 0.0179 and 0.0703 per chicken-day at risk. The four major clinical signs observed were cyanotic combs and wattles, oedema of the head and face, drowsiness and huddling and ecchymotic discoloration of the leg shanks. The first sign (cyanotic combs and wattles) was equally observed in chickens from commercial and backyard farms (p = 0.44) but the second (oedema of the head and face) was more common in backyard farms (p = 0.03).

Observations on the incidence and aetiology of valvular endocarditis in broiler breeders and detection of a newly described taxon of Pasteurellaceae, Avibacterium endocarditidis.

A total of 122 dead broiler breeders randomly selected from a flock showing normal production parameters and covering the age from 44 to 61 weeks were subjected to a comprehensive routine post-mortem examination including examination for lesions of endocarditis. Forty-two hens (34%) showed valvular endocarditis caused by Avibacterium endocarditidis (43%), Enterococcus faecalis (31%), Staphylococcus aureus (5%) and Streptococcus pluranimalium (5%), while growth was not obtained from 17% with the methods used for isolation. Gross lesions associated with the different bacterial pathogens did not allow separation according to pathogens involved. Port of entry and pathogenesis associated with the high prevalence of valvular endocarditis remained speculative. The present findings demonstrated the newly described species of Pasteurellaceae, Avibacterium endocarditidis associated with endocarditis in chickens and confirm previous observations on the prevalence of endocarditis in chickens, partly explaining the slightly increased mortality normally observed in broiler breeders during the last weeks of production.

The space-time clustering of highly pathogenic avian influenza (HPAI) H5N1 outbreaks in Bangladesh.

Bangladesh faced two epidemic waves of highly pathogenic avian influenza (HPAI) H5N1 in two consecutive years. The peaks of the waves were observed in February-July 2007 and January-April 2008, respectively. We examined the spatial and temporal patterns of the 293 outbreaks in 143 subdistricts in 2007 and 2008. Global clustering assessed by K-function was seen at distances 150-300 km between subdistricts. Significant local clusters were detected by space-time scan statistics. In both waves, significant primary clusters of HPAI outbreaks were identified in the central part of the country dominated by commercial production systems and in the northwestern part primarily in backyard production systems. Secondary clusters varied from the northwestern part in 2007 and the southern part in 2008. The findings are highly relevant for the successful planning and implementation of control, prevention and surveillance strategies by highlighting areas where detailed investigations should be initiated.

MHC haplotype and susceptibility to experimental infections (Salmonella Enteritidis, Pasteurella multocida or Ascaridia galli) in a commercial and an indigenous chicken breed.

In three independent experimental infection studies, the susceptibility and course of infection of three pathogens considered of importance in most poultry production systems, Ascaridia galli, Salmonella Enteritidis and Pasteurella multocida were compared in two chicken breeds, the indigenous Vietnamese Ri and the commercial Luong Phuong. Furthermore, the association of the Major Histocompatibility Complex (MHC) with disease-related parameters was evaluated, using alleles of the LEI0258 microsatellite as markers for MHC haplotypes. The Ri chickens were found to be more resistant to A. galli and S. Enteritidis than commercial Luong Phuong chickens. In contrast, the Ri chickens were more susceptible to P. multocida, although production parameters were more affected in the Luong Phuong chickens. Furthermore, it was shown that the individual variations observed in response to the infections were influenced by the MHC. Using marker alleles of the microsatellite LEI0258, which is located within the MHC region, several MHC haplotypes were identified as being associated with infection intensity of A. galli. An association of the MHC with the specific antibody response to S. Enteritidis was also found where four MHC haplotypes were shown to be associated with high specific antibody response. Finally, one MHC haplotype was identified as being associated with pathological lesions and mortality in the P. multocida experiment. Although not statistically significant, our analysis suggested that this haplotype might be associated with resistance. These results demonstrate the presence of local genetic resources in Vietnamese chickens, which could be utilized in breeding programmes aiming at improving disease resistance.

Causes of mortality in commercial organic layers in Denmark.

A longitudinal study investigated the courses of mortality in commercial free-range organic layer flocks in Denmark. In total, 15 organic egg-producing flocks from 11 farms were randomly selected among 80 farms registered in Denmark. Four farms with confined egg production on deep litter were included for comparison. Flock sizes ranged from 2260 to 5940 layers. The flocks were monitored from introduction to the layer farm until slaughter. Flock mortalities ranged from approximately 2% to 91%, with a mean of 20.8% for organic flocks compared with 7% for confined flocks on deep litter. In total, 4608 layers were submitted for postmortem examination, representing > 40% of all the dead layers in the investigated flocks. Outbreaks of erysipelas (Erysipelothrix rhusiopathiae) and fowl cholera (Pasteurella multocida) were observed in two and three organic flocks, respectively. The mortality rate reached 91% in one organic flock dually affected by erysipelas and fowl cholera. In six organic flocks, outbreaks of blackhead were diagnosed. Concurrent infections of erysipelas and blackhead were diagnosed in one organic flock. Escherichia coli infections in the form of septicemia were identified in all organic flocks. In addition, cannibalism and constipation contributed significantly to the mortality in some organic flocks. In the confined deep litter flocks, E. coli infection, constipation, and cannibalism represented the most common causes of mortality.

Mannan-binding lectin (MBL) in two chicken breeds and the correlation with experimental Pasteurella multocida infection.

The present study is the first demonstration of an association of the genetic serum Mannan-binding lectin (MBL) concentration with bacterial infections in chickens. The genetic serum MBL concentration was determined in two chicken breeds, and the association with the specific Pasteurella multocida humoral immune response during an experimental infection was examined. Furthermore, we examined the association of the genetic serum MBL concentration with systemic infection. The chickens with systemic infection had a statistically significant lower mean serum MBL concentration than the rest of the chickens, suggesting that MBL plays an important role against P. multocida. A statistically significant negative correlation was found between the specific antibody response and the genetic serum MBL concentration for both breeds. This indicates that MBL in chickens is capable of acting as the first line of defence against P. multocida by diminishing the infection before the adaptive immune response takes over.

Risk factors for infection with highly pathogenic influenza A virus (H5N1) in commercial chickens in Bangladesh.

A matched case-control study was carried out to identify risk factors for highly pathogenic avian influenza A virus (subtype H5N1) infection in commercial chickens in Bangladesh. A total of 33 commercial farms diagnosed with H5N1 before September 9, 2007, were enrolled as cases, and 99 geographically matched unaffected farms were enrolled as control farms. Farm data were collected using a pretested questionnaire, and analysed by matched-pair analysis and multivariate conditional logistic regression. Two factors independently and positively associated with H5N1 infection remained in the final model. They were 'farm accessible to feral and wild animals' (odds ratio [OR] 5.71, 95 per cent confidence interval [CI] 1.81 to 18.0, P=0.003) and 'footbath at entry to farm/shed' (OR 4.93, 95 per cent CI 1.61 to 15.1, P=0.005). The use of a designated vehicle for sending eggs to a vendor or market appeared to be a protective factor (OR 0.14, 95 per cent CI 0.02 to 0.88, P=0.036).

Association of Streptococcus pluranimalium with valvular endocarditis and septicaemia in adult broiler parents.

The genus Streptococcus consists of more than 60 species, but only Streptococcus equi subspecies zooepidemicus, Streptococcus gallolyticus ssp. gallolyticus, Streptococcus gallinaceus, Streptococcus dysgalactiae, Streptococcus mutans and Streptococcus suis have been isolated from poultry. During investigations of the aetiology of increased mortality in broiler parent stock at the end of production, pure cultures of streptococcal-like organisms that could not be classified among these six species were obtained from 24 cases of septicaemia or valvular endocarditis and septicaemia. Phenotypic characterization using the API20 STREP kit identified the isolates as Aerococcus viridans (10), Aerococcus urinae (2), Leuconostoc species (4), Streptococcus salivarius (2), Streptococcus bovis II 3 (1), Enterococcus avium (3), Enterococcus faecium (1) or Gemella morbillorum (1). However, this identification was misleading as subsequent genetic investigations using pulse field gel electrophoresis and sequencing of 16S rRNA genes showed that 19 isolates were classified as Streptococcus pluranimalium, while the remaining isolates were E. avium (3), E. faecium (1) or Lactobacillus species (1). Misidentification by API20 STREP was related to the database provided by the manufacturer, as the phenotypic characteristics could identify these organisms as S. pluranimalium. The isolates of S. pluranimalium belonged to at least three different clones as determined by pulsed field gel electrophoresis of SmaI-digested genomic DNA. The capacity that these isolates had to colonize the valvular endothelium was suggested by the occurrence of valvular endocarditis in 12 of 19 cases. Demonstration of the same clone in all four houses on a farm suggested the pathogenic potential of this organism.

CCR5 and CXCR3 are dispensable for liver infiltration, but CCR5 protects against virus-induced T-cell-mediated hepatic steatosis.

CCR5 and CXCR3 are important molecules in regulating the migration of activated lymphocytes. Thus, the majority of tissue-infiltrating T cells found in the context of autoimmune conditions and viral infections express CCR5 and CXCR3, and the principal chemokine ligands are expressed within inflamed tissues. Accordingly, intervention studies have pointed to nonredundant roles of these receptors in models of allograft rejection, viral infection, and autoimmunity. In spite of this, considerable controversy exists, with many studies failing to support a role for CCR5 or CXCR3 in disease pathogenesis. One possible explanation is that different chemokine receptors may take over in the absence of any individual receptor, thus rendering individual receptors redundant. We have attempted to address this issue by analyzing CCR5(-/-), CXCR3(-/-), and CCR5/CXCR3(-/-) mice with regard to virus-induced liver inflammation, generation and recruitment of effector cells, virus control, and immunopathology. Our results indicate that CCR5 and CXCR3 are largely dispensable for tissue infiltration and virus control. In contrast, the T-cell response is accelerated in CCR5(-/-) and CCR5/CXCR3(-/-) mice and the absence of CCR5 is associated with the induction of CD8(+) T-cell-mediated immunopathology consisting of marked hepatic microvesicular steatosis.