Elucidation of Schistosoma japonicum population dynamics in pigs using PCR-based identification of individuals representing distinct cohorts

The study reported here investigated the interactions of successive infections and acquired resistance of pigs to challenge infections of Schistosoma japonicum. Two morphologically indistinguishable geographical isolates from China (from Anhui and Zhejiang provinces) were used for the infections. The worms of the two isolates were distinguishable by PCR-linked restriction fragment length polymorphism analysis of the nicotinamide adenine dinucleotide phosphate dehydrogenase I gene of the mitochondrial genome. Thirty-two pigs divided into seven groups were used in the experiment. Two groups received a single infection by either the Anhui or the Zhejiang isolate. In Challenge Groups 1, 4, 6, 8 and 12, a primary infection of the Zhejiang isolate was followed by a challenge infection with the Anhui isolate at week 1, 4, 6, 8 or 12 after the primary infection. In this way it was possible to determine whether worms recovered by perfusion originated from the primary or the challenge infection. Only the challenge infection at week 1 resulted in a higher worm burden when compared with a single primary infection with the Zhejiang isolate. The results showed that challenge worms were able to establish, and that the proportion of worms originating from challenge infection increased at the later challenge infections, however without an increase in the total number of worms. In addition, mixed pairs of the two isolates were found in all challenge-infected groups. The results indicate that pigs are able to mount a partial resistance against re-infection with S. japonicum by 4 weeks after a primary infection, but that worms of the challenge infections eventually replace the primary infection. The finding of mixed pairs of the two isolates indicates that worms of S. japonicum are either polygamous or able to wait in solitude for up to 12 weeks for a partner.     

Elucidation of Schistosoma japonicum population dynamics in pigs using PCR-based identification of individuals representing distinct cohorts

The study reported here investigated the interactions of successive infections and acquired resistance of pigs to challenge infections of Schistosoma japonicum. Two morphologically indistinguishable geographical isolates from China (from Anhui and Zhejiang provinces) were used for the infections. The worms of the two isolates were distinguishable by PCR-linked restriction fragment length polymorphism analysis of the nicotinamide adenine dinucleotide phosphate dehydrogenase I gene of the mitochondrial genome. Thirty-two pigs divided into seven groups were used in the experiment. Two groups received a single infection by either the Anhui or the Zhejiang isolate. In Challenge Groups 1, 4, 6, 8 and 12, a primary infection of the Zhejiang isolate was followed by a challenge infection with the Anhui isolate at week 1, 4, 6, 8 or 12 after the primary infection. In this way it was possible to determine whether worms recovered by perfusion originated from the primary or the challenge infection. Only the challenge infection at week 1 resulted in a higher worm burden when compared with a single primary infection with the Zhejiang isolate. The results showed that challenge worms were able to establish, and that the proportion of worms originating from challenge infection increased at the later challenge infections, however without an increase in the total number of worms. In addition, mixed pairs of the two isolates were found in all challenge-infected groups. The results indicate that pigs are able to mount a partial resistance against re-infection with S. japonicum by 4 weeks after a primary infection, but that worms of the challenge infections eventually replace the primary infection. The finding of mixed pairs of the two isolates indicates that worms of S. japonicum are either polygamous or able to wait in solitude for up to 12 weeks for a partner.     

PCR-based identification of individuals of Schistosoma japonicum representing different subpopulations using a genetic marker in mitochondrial DNA.

A mitochondrial NADH dehydrogenase I gene fragment (NDI) was sequenced for three laboratory maintained isolates of Schistosoma japonicum. Comparison of sequences representing the isolates (originally obtained from the Anhui and Zhejiang provinces of the People's Republic of China, and from the Philippines) revealed inter-isolate sequence variations of 0.2-0.6% and no intra-isolate variation was found. The sequences also indicated that while the amplification products of the Zhejiang and Philippine isolates contained a recognition site for the endonuclease RsaI, there was no such site in the Anhui isolate. This was tested by digesting amplification products from a number of individual worms with RsaI. Then an infection experiment was designed to test the value of this genetic marker for studies of the population biology of S. japonicum in the final host. For this, the two Chinese isolates were used. Three groups of mice (A-C) were exposed firstly to a primary infection and then challenge-infected at weeks 4 and 7 of the experiment. In group A the first infection was done with the Anhui isolate, and the two others with the Zhejiang isolate, thereby providing a specific, detectable cohort. In groups B and C the Anhui isolate was used for the second and third infection. All mice were perfused 5 weeks after the last challenge infection, and the NDI was subsequently amplified from DNA of the perfused worms and digested with RsaI. The digestion revealed that while infection groups A and B contained mixed populations of the Anhui and Zhejiang isolates, only Zhejiang worms were present in group C. We concluded that the absence/presence of the RsaI site in the NDI provides a useful marker for the delineation of cohorts of S. japonicum.     

Cryptosporidium tyzzeri and Cryptosporidium muris originated from wild West-European house mice (Mus musculus domesticus) and East-European house mice (Mus musculus musculus) are non-infectious for pigs

Three and 8 week old pigs were inoculated with Cryptosporidium muris HZ206 (Mus musculus musculus isolate), Cryptosporidium tyzerri CR2090 (M. m. musculus isolate) or C. tyzzeri CR4293 (isolate from a hybrid between Mus musculus domesticus and M. m. musculus) at a dose of 1 × 10(7) oocysts per animal. Inoculated pigs showed no detectable infection and no clinical symptoms of cryptosporidiosis during 30 days post infection (DPI), and no macroscopic changes were detected in the digestive tract following necropsy. Developmental stages were not detected in gastrointestinal tract tissue by histology or PCR throughout the duration of the experiment. The infectivity of isolates was verified on SCID mice, in which oocysts shedding started from 4 to 8 DPI. Based on our findings, it can be concluded that pigs are not susceptible to C. muris or C. tyzzeri infection.     

Role of CD4+ T cells and T-independent mechanisms in acquired resistance to Salmonella typhimurium infection

Two aspects of acquired resistance to Salmonella typhimurium infection in BALB/c mice, i.e., the ability to clear the primary inoculum from the spleen and resistance to a secondary challenge, were studied with the use of mAb against T cell subsets. The ability to clear a temperature-sensitive mutant of S. typhimurium from the spleen (assessed at day 21) was abrogated by in vivo treatment with anti-CD4 mAb. Accelerated bacterial clearance could be adoptively transferred into naive mice. In vitro depletion experiments also showed the role of CD4+ T cells in this phenomenon. Depletion of CD8+ T cells had only a marginal effect. Resistance to reinfection in the late phase of the primary infection (day 50) was markedly depressed by in vivo treatment with anti-CD4 mAb, whereas this was not the case during the early phase (day 14). Furthermore, during the early phase of infection athymic nude mice showed increased nonspecific resistance to reinfection. Taken together these results suggest that T-independent mechanisms play a major role in acquired resistance during the early phase of infection.     

Evaluation of pyrantel-tartrate abbreviated Ascaris suum infections for the development of resistance in young pigs against migrating larvae

Crossbred young pigs were used to test whether abbreviated infections with eggs of Ascaris suum can stimulate the acquisition of resistance to challenge. Weanling pigs from an Ascaris-free colony were kept free of A. suum until they were divided into groups at the age of 7-8 weeks. The experimental animals received pyrantel tartrate during the period when they were being exposed to increasing numbers of infective A. suum eggs and challenged 10 days after the last infective dose. Liver milk-spot counts and larval recoveries from the lungs indicated that the strongest resistance was acquired by the animals that received the drug continuously for 6 weeks while being exposed to six weekly infective egg doses. The data do not suggest any drug-related suppression of the resistance response to A. suum infection.     

Abrogation of resistance to severe mousepox in C57BL/6 mice infected with LP-BM5 murine leukemia viruses.

Strain C57BL/6 (B6) mice infected with LP-BM5 murine leukemia virus (MuLV) develop a disease which combines abnormal lymphoproliferation with profound immunosuppression and has many features in common with human acquired immunodeficiency syndrome induced by HTLV-III/LAV retroviruses. To determine whether this LP-BM5 MuLV infection would affect the innate resistance of B6 mice to a naturally occurring, highly virulent murine pathogen, mice were exposed to ectromelia virus at various times after treatment with LP-BM5 viruses. At week 4 after infection with LP-BM5, mice challenged with ectromelia virus were unable to generate a humoral immune response to this virus, and between weeks 8 and 10 after infection, challenged mice lost the ability to generate an ectromelia virus-specific cytotoxic-T-cell response. Loss of the cellular immune responses to ectromelia virus was associated with an increased susceptibility to the lethal effects of the virus.     

Genomic sequence and virulence evaluation of MN184A‐like porcine reproductive and respiratory syndrome virus in Japan

In this study, a porcine reproductive and respiratory syndrome virus (PRRSV) that was isolated from a 9‐week‐old diseased pig on a farm in Japan with a high mortality rate during 2007–2008 was characterized. This unique isolate, designated as Jpn5‐37, did not have a high nucleotide identity in open reading frame 5 against any Japanese isolates. Among all available type 2 PRRSV complete genome sequences, Jpn5‐37 shared the highest nucleotide identity (93.6%) with virulent strain MN184A. The genomic characteristics of Jpn5‐37 were highly conserved with respect to the virulent MN184A, including a continuous eight amino acid deletion in the nonstructural protein 2 region. Moreover, virus distribution, viremia and the gross and microscopic characteristics of lesions were investigated in pigs 10 days post‐inoculation to elucidate the pathogenicity of the isolate. Intranasal inoculation was found to rapidly result in viremia and dissemination of the Jpn5‐37 isolate to several tissues in a similar manner to EDRD1; however, the amounts of Jpn5‐37 RNA in serum were significantly greater. Similarly, the quantities of Jpn5‐37 viral RNA in all organs tested tended to be higher than with EDRD1 infection. Mean rectal temperatures were significantly higher in the Jpn5‐37‐inoculated than in the control group at 4 and 6 days post infection (dpi) and in the EDRD1‐inoculated group at 6 and 8 dpi. These results suggest that the Jpn5‐37 strain replicates and is more efficiently distributed to the organs than is EDRD1 under the same conditions.     

Dot-ELISA assessment of guinea pig antibody responses to repeated Dermacentor andersoni infestations

Acquired resistance to ixodid tick infestation is expressed by cattle and laboratory animals. Humoral factors appear to be involved in host acquired resistance to tick bite; however, specific immune responses have yet to be fully characterized. This study examined tick resistance expressed by Hartley guinea pigs upon repeated infestation with Dermacentor andersoni, and describes longitudinal development of antigen specific immunoglobulin over approximately 180 days. Guinea pigs were infested either 4 times with D. andersoni adults, or twice with nymphs. Both infestation groups, adults and nymphs, demonstrated a significant level of resistance to re-infestation, following initial exposure. Dot enzyme-linked immunosorbent assay (Dot-ELISA) was employed to detect antibody reactive with D. andersoni salivary gland antigens (SGA). Animals infested with adults had antibody that increased at a relatively constant rate until the fourth infestation, which was differentiated by a sharp increase in titer, that was maintained for approximately 2 wk. Guinea pigs that received nymph infestations had SGA-specific antibody; however, titers were lower than those in the adult infestation group. Antibody levels continued to increase approximately 80 days beyond the final (second) infestation for this group. A direct correlation between resistance and antibody titer was not evident, since resistance was relatively stable after the second infestation in both infestation groups, and tick-specific immunoglobulin levels continually increased.     

Fluctuation of antiviral resistance in the intestinal tracts of nude mice infected with a mouse adenovirus

Oral administration of adenovirus strain K87 to BALB/c nude mice resulted in viral proliferation in the intestinal tract up to around week 6 at which point replication was suppressed. In other words, the host acquired resistance. However, this resistance was temporary and the viral infection persisted over a long period with repeated periods of proliferation and resistance. That the appearance of this resistance is the result of infecting mice with the virus and is not due to age difference per se was made clear through experimentation with nude mice of different age groups. However, it was indicated that increase in age is involved in the decreased rate of reproliferation following initial suppression. No evidence of the virus was obtained from any other organ throughout the infection. Furthermore, throughout the persistent infection, even during the aforementioned periods of resistance, no neutralizing antibody was detected from sera, intestinal wall or intestinal content. When spleen cells from BALB/c heterozygous littermate mice was transferred to the nude mice, an earlier onset of antiviral resistance was seen than in nude mice without the transfer, and this was accompanied by a rise in neutralizing antibody titer. From these results, it is believed that the resistance characteristic of nude mice infected by mouse adenovirus is dependent on some factor other than the neutralizing antibody invoked resistance exhibited by euthymic mice.     

The mouse as a suitable host for an isolate of Schistosoma haematobium from Niger.

The host-parasite relationships of a Schistosoma haematobium isolate, originating from Niger, and the white mouse are described. Swiss OF1 albino mice were exposed individually to 200 cercariae and worms were recovered 9, 12, 16 and 20 weeks post infection. The mean worm returns ranged between 10.54 and 13.05% and did not alter significantly between 9 and 20 weeks post infection. The sex ratio of worms was always in favour of males; from 7.09:1 at 9 weeks after infection it decreased regularly to 3.28:1 at 20 weeks. Male worms reached a mean length of 8.72 mm at 20 weeks. From the 12th week post infection, a high number of eggs was found in the liver and gut. At 20 weeks, eggs were also found in the bladder. Viable eggs and infective miracidia were obtained. The infection of Bulinus truncatus from Niger succeeded with a mean rate of 61% after the first passage through mice. The isolate of S. haematobium was maintained in the laboratory during 3 successive passages through mice. These entirely new results are very probably linked to genetic characteristics peculiar to the S. haematobium populations from Niger.     

Role of major histocompatibility complex class II in resistance of mice to naturally acquired infection with Syphacia obvelata

Genetics plays a substantial role in host resistance in many host-parasite interactions. We examined the prevalence of naturally acquired infection with Syphacia obvelata in a number of mouse strains housed in a non-barrier facility. These mice, which included cross-bred and congenic, inbred strains on various genetic backgrounds, differ in the loci for the immune function genes--major histocompatibility complex class II (MHCII), toll-like receptor 4 (Tlr4), and solute carrier family 11, member 1 (Slc11a1)--which allowed comparisons of the impact of these genes on resistance to pinworm infection. Male and female mice of various ages were sampled over an 18-month period; infection was determined by use of the cellophane tape test. Results indicated that mice that were MHCII+/+ had a significantly lower prevalence of infection than did mice that were MHCII-/-. Differences were not seen between male and female mice. Although MHCII+/+ mice had an age-associated decrease in infection prevalence, such decrease was not seen in MHCII-/- mice. In contrast, infection prevalence in mice with the normal Tlr4 gene (Tlr4(LPS-n/LPS-n)) gene did not differ significantly compared with that in mice that were homozygous for either the point mutation (Tlr4(LPS-d/LPS-d)) or deletion (Tlr4(LPS-del/LPS-del)) of that gene. Likewise, the presence (Sle11a1r/r) or absence (Slc11a1s/s) of functional alleles for Slc11a1 had no effect on the prevalence of infection with S. obvelata. In conclusion, presence of MHCII, but not Tlr4 or Slc11a1 significantly influences prevalence of naturally acquired infection with S. obvelata. These data justify further comprehensive analyses of the immune components that are involved in pinworm resistance.     

Recovery of Schistosoma japonicum from Experimentally Infected Pigs by Perfusion of Liver and Mesenteric Veins

An optimized procedure for perfusion of pigs infected with Schistosoma japonicum was developed. The technique involves insertion of a perfusion influx tube into the thoracic descending aorta, clamping vessels to parts of the body which did not need to be perfused (the kidneys, hind legs, etc.) and placing a collection tube directly into the portal vein. In addition, the clamping technique allows for separate perfusion of the liver and intestinal veins. The perfusion medium was a sodium citrate buffer (40°C) to which the vasodilator sodium nitroprusside was added. Furthermore, an experiment was conducted to investigate if the perfusion efficiency, measured by total worm recovery, could be increased if praziquantel was administered prior to perfusion. Twelve pigs were each infected with 1 000 S. japonicum cercariae and their schistosomes were collected 11 weeks later by separate perfusion of the liver and intestinal veins. Six of these pigs were treated orally with praziquantel one hour before perfusion. In general, the vessels of the livers and intestines of all pigs were well perfused, judging by the resulting pale colour of the tissues. Worms from praziquantel treated pigs were collected within 5 min of perfusion as opposed to approximately 20 min in the non-treated pigs. More worms were collected from the livers of the praziquantel treated pigs, indicating a hepatic shift of schistosomes from the intestinal mesenteries. However, comparable numbers of worms were retained in the mesenteric veins following perfusion in the 2 groups, indicating that manual recovery of schistosomes from the intestinal mesenteries is necessary in addition to perfusion for obtaining the total worm counts. Another experiment was conducted to determine if the intensity and/or duration of infection had an effect on the number of worms collected by the perfusion technique. Seventy-two pigs were allocated into 3 groups of 24 pigs each, which were infected with either 100, 500 or 2 000 cercariae per pig. The 3 groups were further divided into 4 subgroups of 6 pigs each which were perfused with our selective technique at 4, 11, 17 or 24 weeks post infection, respectively. All of the pigs received an oral praziquantel treatment prior to perfusion. The results indicated that increasing intensities and/or duration of infection resulted in trapping of schistosomes in intravascular inflammatory reactions which made it more difficult to collect the adult schistosomes by perfusion.     

Differential Immune Response Associated to Malaria Outcome Is Detectable in Peripheral Blood following Plasmodium yoelii Infection in Mice

Malaria infection in humans elicits a wide range of immune responses that can be detected in peripheral blood, but we lack detailed long-term follow-up data on the primary and subsequent infections that lead to naturally acquired immunity. Studies on antimalarial immune responses in mice have been based on models yielding homogenous infection profiles. Here, we present a mouse model in which a heterogeneous course of Plasmodium yoelii lethal malaria infection is produced in a non-congenic ICR strain to allow comparison among different immunological and clinical outcomes. Three different disease courses were observed ranging from a fatal outcome, either early or late, to a self-resolved infection that conferred long-term immunity against re-infection. Qualitative and quantitative changes produced in leukocyte subpopulations and cytokine profiles detected in peripheral blood during the first week of infection revealed that monocytes, dendritic cells and immature B cells were the main cell subsets present in highly-parasitized mice dying in the first week after infection. Besides, CD4⁺CD25^high T cells expanded at an earlier time point in early deceased mice than in surviving mice and expressed higher levels of intracellular Foxp3 protein. In contrast, survivors showed a limited increase of cytokines release and stable circulating innate cells. From the second week of infection, mice that would die or survive showed similar immune profiles, although CD4⁺CD25^high T cells number increased earlier in mice with the worst prognosis. In surviving mice the expansion of activated circulating T cell and switched-class B cells with a long-term protective humoral response from the second infection week is remarkable. Our results demonstrate that the follow-up studies of immunological blood parameters during a malaria infection can offer information about the course of the pathological process and the immune response.     

Depletion of eosinophils by anti-IL-5 monoclonal antibody treatment of mice infected with Trichinella spiralis does not alter parasite burden or immunologic resistance to reinfection.

Mechanisms of parasite killing by eosinophils are widely studied and are often implicated in mediating resistance to parasitic infection, especially in conjunction with specific antibodies. Evidence for the eosinophil as an anti-parasite killer cell in vivo is limited and may not justify the belief that eosinophils engage and/or kill infective helminths. We reexamined this question in a mouse model of trichinosis in which antisera to eosinophils were previously used to show the requirement for eosinophils in resistance to this nematode. The current studies used mAb to IL-5 to suppress eosinophil levels in CF1 mice infected with Trichinella spiralis. In mice given a primary infection and injected with an isotype control mAb or left untreated, the medullary and peripheral blood eosinophil numbers peaked at 3 wk postinfection (PI) and returned to baseline levels by 4 wk PI. Peripheral blood eosinophil numbers in infected mice injected with anti-IL-5 were maintained at levels below those of uninfected normal mice through 4 wk of infection. Histologically, there was a prominent eosinophil accumulation in infected, untreated, or control-mAb-treated mice associated with nurse cell complexes containing infective juveniles in skeletal muscle at 3 and 4 wk PI. This was largely eliminated in mice treated with anti-IL-5 mAb. However, the number of muscle stage juvenile worms recovered 3 and 4 wk PI after acid pepsin digestion was unaffected by eosinophil depletion. Challenge infections, in which mice were infected at day 0 with 125 muscle stage worms and challenged at day 28 PI with 350 muscle stage worms, developed peak eosinophil numbers in bone marrow and peripheral blood 3 wk after primary infection and 2 wk after challenge infection in mice receiving either no treatment or control mAb. In challenged mice receiving anti-IL-5 mAb, medullary and peripheral blood eosinophil numbers remained at or below those of uninfected animals. Although all groups exhibited significant resistance measured as muscle stage worm burdens 56 days PI, eosinophil depletion did not affect resistance of muscle worm recovery. These results suggest that eosinophils are not essential in the control of T. spiralis in either primary or challenge infections of CF1 mice. This in vivo study illustrates the questionable value of in vitro killing assays to assign effector function to any single inflammatory cell type.     

Concomitant and protective immunity in mice exposed to repeated infections with Echinostoma malayanum

Concomitant immunity and its consequence against infection play roles in regulating worm burdens in helminthiasis. Under natural conditions, this immunity is generated by exposure to repeated low dose or trickle infection. In this study, concomitant immunity was induced in mice exposed repeatedly to infection with Echinostoma malayanum and its protective effect on a challenge infection evaluated. A profile of worm burden from exposure to 10 metacercariae/mouse/week rose rapidly during the first 2 weeks reaching a plateau from week 3 to 8 post infection. Based on a cumulative dose of infection, worm recoveries were around 75% in the first 2 weeks, dropped to 50% at week 3 and 19% at week 8. After week 2, adult worm burden was constant and no juvenile worms were found after week 3 of the experiment. To examine the effect of resistance against reinfection, mice in the experimental group were primarily infected with 10 metacercariae/week for 5 weeks, treated with praziquantel and were challenged with 75 metacercariae/animal. The number of worms recovered from the experimental groups was significantly lower than that from naïve control groups beginning from 24 h to 28 days post challenge. The worms in the experimental group showed growth retardation and the proportion of adult worms was lower than that in the control animals especially during the first 3 weeks of the experiment. Parasite fecundity was also suppressed compared with that in the control group. The selective effects of protective immunity on establishment, growth, and fecundity of challenged worms affected the population dynamics of E. malayanum which is a similar phenomenon to concomitant immunity in schistosomiasis.     

Trichinella spiralis: major histocompatibility complex-associated elimination of encysted muscle larvae in swine

A heretofore undescribed host-mediated reactivity against encapsulated muscle larvae (ML) of the nematode Trichinella spiralis is reported. Inbred miniature swine (NIH minipigs) of three independent SLA phenotypes, which received a primary oral dose of 300 T. spiralis ML, successfully resisted a secondary infection of 10,000 ML; however, only pigs of the SLAa/a phenotype exhibited an unusual and highly significant reduction in the numbers of encysted ML from the primary infection (P less than 0.0003). This initial anti-encysted ML reactivity was confirmed in subsequent trials by comparing the prechallenge ML burdens with the reduced ML numbers in primary-infected aa pigs after challenge. Analyses of inbred strains of mice, selected for major histocompatibility type and for resistance or susceptibility to infection with T. spiralis, showed no such anti-encysted ML response. Because elimination of encysted T. spiralis ML had been accomplished previously only through selected drug regimens, our demonstration of a nonpharmacological, host-mediated reactivity against this stage of the parasite in swine highlights the importance of MHC genes in regulating disease resistance in a livestock species.     

Acquired resistance to migrating larvae of Ascaris suum in young pigs by repeated drug-abbreviated infections

Cross-bred 3- and 8-wk-old pigs were used to test whether drug-abbreviated infections with Ascaris suum can stimulate acquired resistance to challenge. During the immunization period, both age groups of animals were infected with increasing numbers of A. suum eggs (500, 1,000, 2,000, 5,000, 10,000, and 20,000) at 7-day intervals while the pigs were receiving pyrantel tartrate in the feed. Two days after the last infective dose, animals were placed on unmedicated feed for 8 days and then challenged with 10,000 eggs. All pigs were killed 7 days after challenge, and milk spots on the livers and larvae recovered from the lungs were counted. Larval recoveries from lungs of the immunized animals were significantly smaller than those from the unimmunized animals in both age groups, suggesting that the pigs were capable of acquiring strong resistance to parasitic infections. In immunized animals, challenge infection did not contribute significantly to milk spot formation. The number of milk spots was significantly greater in the older animals, indicating that milk spot formation may be age related.     

Conjugated linoleic acid fails to worsen insulin resistance but induces hepatic steatosis in the presence of leptin in ob/ob mice

Conjugated linoleic acid (CLA) induces insulin resistance preceded by rapid depletion of the adipokines leptin and adiponectin, increased inflammation, and hepatic steatosis in mice. To determine the role of leptin in CLA-mediated insulin resistance and hepatic steatosis, recombinant leptin was coadministered with dietary CLA in ob/ob mice to control leptin levels and to, in effect, negate the leptin depletion effect of CLA. In a 2 x 2 factorial design, 6 week old male ob/ob mice were fed either a control diet or a diet supplemented with CLA and received daily intraperitoneal injections of either leptin or vehicle for 4 weeks. In the absence of leptin, CLA significantly depleted adiponectin and induced insulin resistance, but it did not increase hepatic triglyceride concentrations or adipose inflammation, marked by interleukin-6 and tumor necrosis factor-alpha mRNA expression. Insulin resistance, however, was accompanied by increased macrophage infiltration (F4/80 mRNA) in adipose tissue. In the presence of leptin, CLA depleted adiponectin but did not induce insulin resistance or macrophage infiltration. Despite this, CLA induced hepatic steatosis. In summary, CLA worsened insulin resistance without evidence of inflammation or hepatic steatosis in mice after 4 weeks. In the presence of leptin, CLA failed to worsen insulin resistance but induced hepatic steatosis in ob/ob mice.     

An experimental model of Actinobacillus suis infection in mice

Actinobacillus suis is an opportunistic pathogen of high health status swine and is associated with fatal septicemia, especially in neonatal pigs. A practical model of A. suis is unavailable currently. However, some evidence suggests that A. suis can infect nonporcine species. We therefore hypothesized that a mouse model of A. suis infection might be possible. To test this idea, we challenged CD1 mice with 3 strains of A. suis (2 porcine [SO4 and H91-0380] and 1 feline [96-2247]) by intranasal and intraperitoneal routes. We also evaluated the effects of coadministration of hemoglobin and immunosuppression by dexamethasone on the susceptibility of mice to A. suis infection. The feline and H91-0380 porcine strains induced clinical signs of acute disease and necrotizing pneumonia in mice similar to those seen in pigs. Although few bacteria were recovered, dissemination of A. suis was widespread. Generally, mice infected with the feline A. suis isolate had more severe clinical signs and higher bacterial titers than did mice infected with either of the porcine strains. Pretreatment of the mice with dexamethasone or addition of 2% porcine hemoglobin to the challenge inoculum appeared to hasten the onset of clinical signs by the porcine strains but had no significant effect on moribundity. These experiments demonstrate that mice can be infected with A. suis and subsequently develop pneumonia and bacteremia comparable to that seen in pigs, suggesting that mice may be used as a model for studying infection in swine.