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We assessed the concentrations of metals and other trace elements in two of the most common seabird species breeding on Svalbard, the black-legged kittiwake (Rissa tridactyla) and the Brünnich's guillemot (Uria lomvia). Both of these species feed mostly on fish and crustaceans but have different foraging strategies, kittiwakes being surface feeders while guillemots are divers. We investigated the concentrations of arsenic (As), cadmium (Cd), chromium (Cr), copper (Cu), mercury (Hg), lead (Pb), selenium (Se) and zinc (Zn) in the plasma and body feathers of black-legged kittiwakes (n = 17), as well as in the body feathers of Brünnich's guillemots (n = 13). Samples were collected from adult birds at two time points, one week apart during July 2017 in Kongsfjorden, Svalbard. Of the non-essential trace elements, As was found at the highest median concentration at both the first (56.23 ng/g ww) and second (39.99 ng/g ww) sampling timepoints in the kittiwake plasma. When separating for the sexes, as well as sampling time, males sampled at the first sampling time point had significantly higher concentrations of As (median at 0.087 ng/g versus 0.039 ng/g) and Se (median 0.26 ng/g versus 0.16 ng/g) compared to males sampled at the second time point. There was no significant difference in plasma concentrations between females at first and second sampling time points. Kittiwake feathers contained significantly higher concentrations of As, Cd and Hg than guillemot feathers, while guillemot feathers had significantly higher concentrations of Cu, Pb and Zn. However, of the non-essential elements in both kittiwake and guillemot feathers Hg was found with the highest median concentrations at 5160 and 1080 ng/g, respectively, thus in kittiwakes exceeding the level of 5000 ng/g associated with adverse effect (e.g., impaired reproduction). Levels of Hg and Se found in the kittiwake feathers were higher than previous studies on seabirds in the Arctic., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Anthropogenic pollution is identified as an important threat to bird and other wildlife populations. Many metals and toxic elements, along with poly- and perfluoroalkyl substances (PFASs) are known to induce immunomodulation and have previously been linked to increased pathogen prevalence and infectious disease severity. In this study, the house sparrow (Passer domesticus) was investigated at the coast of Helgeland in northern Norway. This population is commonly infected with the parasitic nematode "gapeworm" (Syngamus trachea), with a prevalence of 40-60 % during summer months. Gapeworm induces severe respiratory disease in birds and has been previously demonstrated to decrease survival and reproductive success in wild house sparrows. The aim of this study was to investigate whether a higher exposure to pollution with PFASs, metals and other elements influences gapeworm infection in wild house sparrows. We conducted PFASs and elemental analysis on whole blood from 52 house sparrows from Helgeland, including analyses of highly toxic metals such as lead (Pb), mercury (Hg) and arsenic (As). In addition, we studied gapeworm infection load by counting the parasite eggs in faeces from each individual. We also studied the expression of microRNA 155 (miR155) as a key regulator in the immune system. Elevated blood concentrations of Pb were found to be associated with an increased prevalence of gapeworm infection in the house sparrow. The expression of miR155 in the plasma of the house sparrow was only weakly associated with Pb. In contrast, we found relatively low PFASs concentrations in the house sparrow blood (∑ PFASs 0.00048-354 μg/L) and PFASs were not associated to miR155 nor infection rate. The current study highlights the potential threat posed by Pb as an immunotoxic pollutant in small songbirds., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Essa Ahsan Khan reports financial support was provided by Norwegian University of Science and Technology. Henrik Jensen and Veerle L.B. Jaspers reports financial support was provided by Research Council of Norway. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Purpose: Reverse total shoulder replacement (rTSR) improves pain and function in patients with a wide range of shoulder pathology. Anterosuperior (AS) and deltopectoral (DP) approaches are widely used to gain access to the glenohumeral joint in arthroplasty. Our aim was to systematically review the literature comparing outcomes of these two approaches when performing rTSR for degenerative glenohumeral arthritis., Methods: Systematic review was performed with an electronic multi-database search (Pubmed, Medline & Embase) according to PRISMA guidelines on 18th September 2022. Data from published studies of any study design that met the inclusion criteria were extracted, reviewed and synthesized., Results: A total of 38 studies were identified for full text review, of which four were included. No significant difference in pain and range of motion were observed between approaches. Scapular notching was more common in the anterosuperior approach. Low rates of instability and intra-operative fracture were observed in both., Conclusion: Both approaches demonstrate similar clinical outcomes with reference to pain, range of motion and complications when performing rTSR indicated for degenerative joint disease. However, further well-designed studies are required., (© 2023. The Author(s), under exclusive licence to Springer-Verlag France SAS, part of Springer Nature.)

Background:  For many years the long head of biceps tendon (LHBT) rupture has been described and is commonly identified by weakness, cramping, and the so-called "Popeye" sign. Traditionally, this was treated non-operatively, likely reflecting patient factors and the technical difficulty in reattaching a degenerative and shortened tendon. In contrast, traumatic distal biceps rupture is now commonly repaired despite historically being managed non-operatively. The advent of a convenient and reproducible surgical technique led to an increase in the rate of fixation, thereby improving the cramping and weakness associated with non-operative treatment. Given recent surgical advances within this field, many techniques are now present for LHBT pathology. We describe results from a cohort of patients suffering traumatic LHBT rupture who sought a surgical solution to improve their symptoms., Methods:  Over four years, 18 male patients underwent surgical intervention for isolated traumatic LHBT rupture. The technique used involved an open subpectoral tenodesis with fixation of the LHBT into the bicipital groove. Postoperative immobilization using a sling was recommended for six weeks prior to a progressive rehabilitation program. Patients were assessed with pre- and postoperative visual analog scores (VAS) for pain and American Shoulder and Elbow Society (ASES) scores., Results: The mean patient age at the time of surgery was 49 years (range: 26-65 years). The mean time to surgery was nine weeks (range: 2-24 weeks). All patients showed an improvement following surgery with a mean pre-op ASES score of 33 (range: 10-60) compared to a post-op score of 92.6 (range: 85-100). All patients were able to return to work and sport, with all but one returning to the same functional demand level of work. The mean pre-op pain VAS was 6.3 (out of 10) compared to 0.2 post-op. All patients had a requirement for analgesia pre-operatively and none had postoperatively. No surgical complications were observed. No correlation was observed between the time to surgery and the outcome., Discussion:  LHBT rupture is often treated non-operatively as few studies within the literature describe the surgical technique and outcomes from surgical intervention. When treated non-operatively, patients complain of pain, cramping, and cosmetic deformity known as the "Popeye" sign. Following a traumatic rupture of the LHBT, we have demonstrated excellent outcomes using a standard approach and common fixation technique that has the potential to improve the functional outcome for symptomatic patients., Conclusion:  Open subpectoral biceps tenodesis is associated with excellent outcomes in symptomatic patients following isolated LHBT rupture., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2022, Waugh et al.)

Old lead-zinc (Pb-Zn) mining sites in Greenland have increased the environmental concentration of Pb in local marine organisms, including the shorthorn sculpin. Organ metal concentrations and histopathology have been used in environmental monitoring programs to evaluate metal exposure and subsequent effects in shorthorn sculpins. So far, no study has reported the impact of heavy metals on gene expression involved in metal-related stress and immune responses in sculpins. The aim of this study was to investigate the effect of exposure to environmentally relevant waterborne Pb (0.73 ± 0.35 μg/L) on hepatic gene expression of metallothionein (mt), immunoglobulin M (igm), and microRNAs (miRNAs; mir132 and mir155) associated with immune responses in the shorthorn sculpin compared to a control group. The mt and igm expression were upregulated in the Pb-exposed group compared to the control group. The transcripts of mir132 and mir155 were not different in sculpins between the Pb-exposed and control group; however, miRNA levels were significantly correlated with Pb liver concentrations. Furthermore, there was a positive correlation between liver Pb concentrations and igm, and a positive relationship between igm and mir155. The results indicate that exposure to Pb similar to those concentrations reported in in marine waters around Greenland Pb-Zn mine sites influences the mt and immune responses in shorthorn sculpins. This is the first study to identify candidate molecular markers in the shorthorn sculpins exposed to waterborne environmentally relevant Pb suggesting mt and igm as potential molecular markers of exposure to be applied in future assessments of the marine environment near Arctic mining sites., (© 2022. The Author(s).)

The validation of the use of primary cell lines from non-lethal matrixes of feathers and blood of nestlings of a wild bird species, the tawny owl (Strix aluco) is described. Tawny Owl Feather Fibroblast (TOFF) cells and peripheral blood mononuclear cells (PBMCs) were isolated and cultured from the pulp of the secondary wing feathers and whole blood respectively from free-living tawny owl nestlings. Cell growth was registered up until 48 h for both the PBMC cells and the TOFFs. The validation of these primary cell lines in free-living birds has the potential to advance the assessment of immunotoxicological effects in wildlife via non-lethal manner. They provide a key tool with which to study cell toxicity and responses to environmental stressors on a cellular level in wild bird species of interest., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Kroglund, Eide, Østnes, Kroglund, Frisli and Waugh.)

Exposure to certain perfluoroalkyl acids (PFAAs) can have considerable effects on the endocrine and immune systems, although such effects remain largely uncharacterized in wildlife. Using an apex avian predator, we investigated possible relationships of thyroid hormones (THs), specifically free (F) and total (T) thyroxine (FT4; TT4) and triiodothyronine (FT3; TT3), and the expression of an immune-related microRNA biomarker (i.e., miR-155), with the concentrations of 11 PFAAs in nestling peregrine falcons (Falco peregrinus). Nestling peregrines (n = 56; usually two chicks of each sex per nest) were blood sampled when 23 ± 4 days old in urban and rural regions of the Laurentian Great Lakes Basin (Ontario, Canada) in 2016 and 2018. The circulating concentrations of several PFAAs were significantly associated with THs and estimated thyroid gland activity (TT3:TT4; FT3:FT4), including PFHxS (FT3; FT3:FT4), PFDS (TT3; TT3:TT4), PFOA (TT4; FT3:FT4), PFTeDA (TT4; FT3:FT4), PFHxDA (TT4; TT3:TT4) and ΣPFCAs (TT4). Our novel evaluation of miR-155 in peregrine nestlings identified significantly negative relationships of plasma miR-155 counts with PFHxS and PFOA concentrations, indicating potential down-regulation of miR-155 expression and impaired immunity. Several PFAA homologues significantly predicted the variation in THs and miR-155 in conjunction with year (e.g., inter-annual differences in weather, ambient temperature, rainfall), region (urban/rural), nestling age, and/or diet (trophic position; δ 15 N), which suggests that multiple environmental and biological stressors, including PFAA exposure, influenced thyroid activity and immune function in these nestlings. Further research is warranted to identify the mechanisms and additional impacts of PFAA-related thyroid and immune disruption on the growth, development, and health risks in developing birds., Competing Interests: Declaration of competing interest The authors declare that there are no intellectual or financial conflicts of interest., (Crown Copyright © 2021. Published by Elsevier B.V. All rights reserved.)

Many persistent organic pollutants, such as polychlorinated biphenyls (PCBs), have high immunomodulating potentials. Exposure to them, in combination with virus infections, has been shown to aggravate outcomes of the infection, leading to increased viral titers and host mortality. Expression of immune-related microRNA (miR) signaling pathways (by host and/or virus) have been shown to be important in determining these outcomes; there is some evidence to suggest pollutants can cause dysregulation of miRNAs. It was thus hypothesized here that modulation of miRNAs (and associated cytokine genes) by pollutants exerts negative effects during viral infections. To test this, an in vitro study on chicken embryo fibroblasts (CEF) exposed to a PCB mixture (Aroclor 1260) and then stimulated with a synthetic RNA virus (poly(I:C)) or infected with a lymphoma-causing DNA virus (Gallid Herpes Virus 2 [GaHV-2]) was conducted. Using quantitative real-time PCR, expression patterns for mir-155 , pro-inflammatory TNF α and IL-8 , transcription factor NF- κ B1 , and anti-inflammatory IL-4 were investigated 8, 12, and 18 h after virus activation. The study showed that Aroclor1260 modulated mir-155 expression, such that a down-regulation of mir-155 in poly(I:C)-treated CEF was seen up to 12 h. Aroclor1260 exposure also increased the mRNA expression of pro-inflammatory genes after 8 h in poly(I:C)-treated cells, but levels in GaHV-2-infected cells were unaffected. In contrast to with Aroclor1260/poly(I:C), Aroclor1260/GaHV-2-infected cells displayed an increase in mir-155 levels after 12 h compared to levels seen with either individual treatment. While after 12 h expression of most evaluated genes was down-regulated (independent of treatment regimen), by 18 h, up-regulation was evident again. In conclusion, this study added evidence that mir-155 signaling represents a sensitive pathway to chemically-induced immunomodulation and indicated that PCBs can modulate highly-regulated innate immune system signaling pathways important in determining host immune response outcomes during viral infections.

Southern hemisphere humpback whales are classified as high-fidelity Antarctic krill consumers and as such are vulnerable to variability and long-term changes in krill biomass. Evidence of heterogeneous feeding patterns of east coast of Australia migrating humpback whales has been observed, warranting a comprehensive assessment of interannual variability in their diet. We examined the lipid and fatty acid profiles of individuals of the east coast of Australia migrating stock sampled between 2008 and 2018. The use of live-sampled blubber biopsies showed that fatty acid profiles varied significantly among all years. The two trophic indicator fatty acids for Antarctic krill, 20:5ω3 and 22:6ω3 remained largely unchanged across the 10-year period, suggesting that Antarctic krill is the principal prey item. A distance-based linear model showed that 33% of the total variation in fatty acid profiles was explained by environmental variables and climate indices. Most of the variation was explained by the Southern Annular Mode (23.7%). The high degree of variability observed in this study was unexpected for a species that is thought to feed primarily on one prey item. We propose that the observed variability likely arises from changes in the diet of Antarctic krill rather than changes in the whale's diet.

Background: Wild aquatic birds serve as the natural reservoir for avian influenza virus (AIV), a disease with significant implications for avian and mammalian health. Climate change is predicted to impact the dynamics of AIV, particularly in areas such as the Arctic, but the baseline data needed to detect these shifts is often unavailable. In this study, plasma from two species of gulls breeding on the high-Arctic Svalbard archipelago were screened for antibodies to AIV., Results: AIV antibodies were found in black-legged kittiwake (Rissa tridactyla) samples from multiple years, as well as in glaucous gulls (Larus hyperboreous) samples., Conclusions: Despite small sample sizes, evidence of exposure to AIV was found among Svalbard gulls. A wider survey of Svalbard avian species is warranted to establish knowledge on the extent of AIV exposure on Svalbard and to determine whether active infections are present.

Background: Since 2016, incursions of highly pathogenic avian influenza virus (HPAIV) H5N8 clade 2.3.4.4b have caused unprecedented clinical signs and mortality in white-tailed eagles (WTE; Haliaeetus albicilla) across Europe and have been found to be infecting other raptor species, such as the northern goshawk (NG; Accipiter gentilis). Before this study, no screening of Norwegian raptors had been undertaken., Results: Plasma samples from 43 white-tailed eagle and 29 northern goshawk nestlings, from several locations across Norway were screened for antibodies to avian influenza viruses. No antibodies, and thus, no evidence of AIV exposure, were found in these Norwegian raptors. No clinical signs of AIV were observed in 43 white tailed eagles and 29 northern goshawks., Conclusions: There are currently no indications that white-tailed eagles and northern goshawks inhabiting Norway are threatened by the recent HPAIV outbreaks in other areas of Europe. Ongoing monitoring should, however, be maintained to detect potential future outbreaks.

Background: Per- and polyfluoroalkyl substances (PFASs) are environmentally persistent and bioaccumulative chemicals. Immunomodulation is among the most concerning of toxic effects linked with PFAS exposure in mammalian models. However, no studies had yet shown this to be true in birds. Thus, we designed and conducted the first study to determine if PFASs could cause immunomodulation in birds. Secondly, we wanted to determine the effects on an avian host when exposed not only to immunomodulating chemicals, but also to a viral challenge. The aim, to determine if PFAS mediated immunmodulation functionally affects a pathogen challenge for a host. As innate immune system signalling pathways initiate crucial responses against a pathogen challenge, and are lesser studied than their adaptive counterparts, we focused on these pathways. To provide the first information on this, an in vitro experiment was designed and performed using chicken embryo fibroblasts exposed to perfluorooctane sulfonate (PFOS) (22 ppm) and immune markers characterised before and after being infected with gallid herpesvirus-2 (GaHV-2)., Results: The expression of two pro-inflammatory cytokines, namely interleukin 8 (IL-8) and tumor necrosis factor alpha (TNF-α), the nuclear factor 'kappa-light-chain-enhancer' of activated B-cells (NF-κB), and the anti-inflammatory cytokine interleukin 4 (IL-4) were investigated in various scenarios. These results showed that exposure to PFOS decreased immune gene expression in chicken fibroblasts from 36 h post-exposure. Next, it was shown that this decrease could be mitigated by infection with gallid herpesvirus-2, which increased gene expression back to the baseline/control levels., Conclusions: Not only is this the first study to provide the expected evidence that PFOS has immunomodulatory potential in birds, it also provides unexpected data that virus infections can mitigate this negative effect. Thereby, further research, including in vivo and in situ studies, on the impact of PFOS on host-virus interactions is now warranted, as it has been overlooked and might contribute to our understanding of recent disease outbreaks in wildlife. The mechanisms by which gallid herpesvirus mitigates immunomodulation were beyond the scope of this study, but are now of interest for future study.

Chlamydia pecorum is responsible for causing ocular infection and disease which can lead to blindness in koalas (Phascolarctos cinereus). Antibiotics are the current treatment for chlamydial infection and disease in koalas, however, they can be detrimental for the koala's gastrointestinal tract microbiota and in severe cases, can lead to dysbiosis and death. In this study, we evaluated the therapeutic effects provided by a recombinant chlamydial major outer membrane protein (MOMP) vaccine on ocular disease in koalas. Koalas with ocular disease (unilateral or bilateral) were vaccinated and assessed for six weeks, evaluating any changes to the conjunctival tissue and discharge. Samples were collected pre- and post-vaccination to evaluate both humoral and cell-mediated immune responses. We further assessed the infecting C. pecorum genotype, host MHC class II alleles and presence of koala retrovirus type (KoRV-B). Our results clearly showed an improvement in the clinical ocular disease state of all seven koalas, post-vaccination. We observed increases in ocular mucosal IgA antibodies to whole C. pecorum elementary bodies, post-vaccination. We found that systemic cell-mediated immune responses to interferon-γ, interleukin-6 and interleukin-17A were not significantly predictive of ocular disease in koalas. Interestingly, one koala did not have as positive a clinical response (in one eye primarily) and this koala was infected with a C. pecorum genotype (E') that was not used as part of the vaccine formula (MOMP genotypes A, F and G). The predominant MHC class II alleles identified were DAb*19, DAb*21 and DBb*05, with no two koalas identified with the same genetic sequence. Additionally, KoRV-B, which is associated with chlamydial disease outcome, was identified in two (29%) ocular diseased koalas, which still produced vaccine-induced immune responses and clinical ocular improvements post-vaccination. Our findings show promise for the use of a recombinant chlamydial MOMP vaccine for the therapeutic treatment of ocular disease in koalas., Competing Interests: The authors have declared that no competing interests exist.

Chlamydia pecorum is a mucosal infection, which causes debilitating disease of the urinary tract, reproductive tract and ocular sites of koalas (Phascolarctos cinereus). While antibiotics are available for treatment, they are detrimental to the koalas' gastrointestinal tract microflora leaving the implementation of a vaccine as an ideal option for the long-term management of koala populations. We have previously reported on the successes of an anti-chlamydial recombinant major outer membrane protein (rMOMP) vaccine however, recombinant protein based vaccines are not ideal candidates for scale up from the research level to small-medium production level for wider usage. Peptide based vaccines are a promising area for vaccine development, because peptides are stable, cost effective and easily produced. In this current study, we assessed, for the first time, the immune responses to a synthetic peptide based anti-chlamydial vaccine in koalas. Five healthy male koalas were vaccinated with two synthetic peptides derived from C. pecorum MOMP and another five healthy male koalas were vaccinated with full length recombinant C. pecorum MOMP (genotype G). Systemic (IgG) and mucosal (IgA) antibodies were quantified and pre-vaccination levels compared to post-vaccination levels (12 and 26 weeks). MOMP-peptide vaccinated koalas produced Chlamydia-specific IgG and IgA antibodies, which were able to recognise not only the genotype used in the vaccination, but also MOMPs from several other koala C. pecorum genotypes. In addition, IgA antibodies induced at the ocular site not only recognised recombinant MOMP protein but also, whole native chlamydial elementary bodies. Interestingly, some MOMP-peptide vaccinated koalas showed a stronger and more sustained vaccine-induced mucosal IgA antibody response than observed in MOMP-protein vaccinated koalas. These results demonstrate that a synthetic MOMP peptide based vaccine is capable of inducing a Chlamydia-specific antibody response in koalas and is a promising candidate for future vaccine development., Competing Interests: The authors have declared that no competing interests exist.

Southern hemisphere humpback whales (Megaptera novaeangliae) rely on summer prey abundance of Antarctic krill (Euphausia superba) to fuel one of the longest-known mammalian migrations on the planet. It is hypothesized that this species, already adapted to endure metabolic extremes, will be one of the first Antarctic consumers to show measurable physiological change in response to fluctuating prey availability in a changing climate; and as such, a powerful sentinel candidate for the Antarctic sea-ice ecosystem. Here, we targeted the sentinel parameters of humpback whale adiposity and diet, using novel, as well as established, chemical and biochemical markers, and assembled a time trend spanning 8 years. We show the synchronous, inter-annual oscillation of two measures of humpback whale adiposity with Southern Ocean environmental variables and climate indices. Furthermore, bulk stable isotope signatures provide clear indication of dietary compensation strategies, or a lower trophic level isotopic change, following years indicated as leaner years for the whales. The observed synchronicity of humpback whale adiposity and dietary markers, with climate patterns in the Southern Ocean, lends strength to the role of humpback whales as powerful Antarctic sea-ice ecosystem sentinels. The work carries significant potential to reform current ecosystem surveillance in the Antarctic region., (© 2017 John Wiley & Sons Ltd.)

Polychlorinated biphenyls (PCBs), and similar environmental contaminants, have been linked to virus outbreaks and increased viral induced mortality since the 1970s. Yet the mechanisms behind this increased susceptibility remain elusive. It has recently been illustrated that the innate immune viral detection system is tightly regulated by small non-coding RNAs, including microRNAs (miRNAs). For virus infections miRNA-155 expression is an important host response against infection, and deregulation of this miRNA is closely associated with adverse outcomes. Thus, we designed a targeted in vitro study using primary chicken fibroblasts, first exposed to a mixture of PCBs (Arochlor-1250) before being stimulated with a synthetic RNA virus (poly I:C), to determine if PCBs have the potential to deregulate miRNA-155. In this paper, we provide the first data for the deregulation of miRNA-155 when a host is exposed to a mixture of PCBs before a virus infection. Thus, we provide important evidence that PCBs can be involved in the deregulation of important miRNA pathways involved in the immune system; thereby demonstrating novel insights into the mechanism of PCB toxicity on the immune system., (© 2018 APMIS. Published by John Wiley & Sons Ltd.)

Chlamydial disease continues to be one of the main factors threatening the long-term survival of the koala (Phascolarctos cinereus). Despite this, large epidemiological studies of chlamydial infection and disease in wild koala populations are lacking. A better understanding of the prevalence, transmission and pathogenesis is needed to improve control measures, such as the development of vaccines. We investigated the prevalence of Chlamydia pecorum infection and disease in 160 koalas in a peri-urban wild population in Queensland, Australia and found that 31% of koalas were Chlamydia PCR positive and 28% had clinically detectable chlamydial disease. Most infections were at the urogenital site (27%; both males and females) with only 14% at the ocular site. Interestingly, we found that 27% (4/15) of koalas considered to be sexually immature (9-13 months) were already infected with C. pecorum, suggesting that a significant percentage of animals are infected directly from their mother. Ocular infection levels were less prevalent with increasing age (8% in koalas older than 4 years), whereas the prevalence of urogenital tract infections remained high into older age (26% in koalas older than 4 years), suggesting that, after mother-to-young transmission, C. pecorum is predominantly a sexually transmitted infection. While 28% of koalas in this population had clinically detectable chlamydial disease (primarily urogenital tract disease), many PCR positive koalas had no detectable disease and importantly, not all diseased animals were PCR positive. We also observed higher chlamydial loads in koalas who were C. pecorum infected without clinical disease than in koalas who were C. pecorum infected with clinical disease. These results shed light on the potential mechanisms of transmission of C. pecorum in koalas and also guide future control measures, such as vaccination.

Introduction: Chlamydia trachomatis is the etiological agent for the most prevalent bacterial sexually transmitted infection in both developed and developing countries. The aim of present study was to characterize the antibody response between two groups of individuals, having either a single C. trachomatis infection and or repeated infections., Material and Methods: Current study consisted of two groups, one with an initial Chlamydia infection and a second with repeated infections. A titre based estimation of specific serum (IgG and IgA) levels using ELISA were performed, which further validated by western blot. In vitro neutralizing ability of each patient's serum against both homologous and heterologous strains was also determined., Results: Individuals infected with one of the C. trachomatis serovars D, E or K exhibited a strong systemic antibody response as characterized by ELISA and western blot. These individuals may have developed at least some level of protection as they only represented single infection. By comparison, individuals infected with serovar D, E or F that exhibited low systemic antibody response often presented repeated C. trachomatis infections, suggesting an association with poor immune response. An in vitro neutralizing level of 60-90% was observed in the human sera against homologous serovar D and two heterologous C. trachomatis serovars E and K, compared to <40% against heterologous serovars F., Conclusion: Individuals infected with serovars D and K showed a potential association between circulating antibody response and re-infection risk. While the patients infected with serovars E showed a disconnection between systemic antibody response and re-infection risk.

The virulence of chlamydial infection in wild koalas is highly variable between individuals. Some koalas can be infected (PCR positive) with Chlamydia for long periods but remain asymptomatic, whereas others develop clinical disease. Chlamydia in the koala has traditionally been studied without regard to coinfection with other pathogens, although koalas are usually subject to infection with koala retrovirus (KoRV). Retroviruses can be immunosuppressive, and there is evidence of an immunosuppressive effect of KoRV in vitro. Originally thought to be a single endogenous strain, a new, potentially more virulent exogenous variant (KoRV-B) was recently reported. We hypothesized that KoRV-B might significantly alter chlamydial disease outcomes in koalas, presumably via immunosuppression. By studying sub-groups of Chlamydia and KoRV infected koalas in the wild, we found that neither total KoRV load (either viraemia or proviral copies per genome), nor chlamydial infection level or strain type, was significantly associated with chlamydial disease risk. However, PCR positivity with KoRV-B was significantly associated with chlamydial disease in koalas (p = 0.02961). This represents an example of a recently evolved virus variant that may be predisposing its host (the koala) to overt clinical disease when co-infected with an otherwise asymptomatic bacterial pathogen (Chlamydia).

The infection status of angiostrongylosis in Jamaica was assessed in wild rats and molluscs in the 5 years following the major outbreak of eosinophilic meningitis (EM) in 2000. Parasitological analyses of 297 Rattus rattus and 140 Rattus norvegicus, and 777 terrestrial molluscs from all 14 Parishes on the island revealed Angiostrongylus cantonensis in 32·0% of the rats and in 12·5% of the molluscs. Multivariate analyses confirmed that A. cantonensis occurred significantly more frequently in R. rattus (Odds Ratio [OR] = 1·76), while mean infection intensity in R. rattus was also significantly higher (16·8) than R. norvegicus (11·3) (Mann-Whitney U-test: P = 0·01). Third-stage larvae of A. cantonensis were detected in 29% of 86 Pleurodonte spp.; in 20% of five Poteria spp.; in 18·7% of 369 Thelidomus asper; in 11% of 18 Sagda spp.; and in 6% of 24 veronicellid slugs. Most rodent infections occurred in Northeastern Jamaica (OR = 11·66), a region where infected molluscs were also abundant. Given the prevalence of A. cantonensis infection in rats has significantly increased since the 2000 outbreak, and that a survey of human infections revealed at least ten autochthonous cases in the last 15 years, angiostrongylosis persists as an important zoonosis in Jamaica.

Chlamydia pecorum infections are debilitating in the koala, contributing significantly to morbidity and mortality, with current antibiotic treatments having minimal success and adversely affecting gut microflora. This, combined with the sometimes-asymptomatic nature of the infection, suggests that an efficacious anti-chlamydial vaccine is required to control chlamydial infections in the koala. To date vaccination studies have focused primarily on female koalas, however, given the physiological differences between male and female reproductive tracts, we tested the efficacy of a vaccine in 12 captive male koalas. We evaluated the potential of both subcutaneous and intranasal vaccine delivery to elicit mucosal immunity in male koalas. Our results showed that both intranasal and subcutaneous delivery of a vaccine consisting of C. pecorum major outer membrane protein (MOMP) and the adjuvant immunostimulating complex (ISC) induced significant immune responses in male koalas. Subcutaneous immunization elicited stronger cell-mediated responses in peripheral blood lymphocytes (PBL), and greater plasma antibody levels whereas the intranasal immunization elicited stronger humoral responses in urogenital tract (UGT) secretions. This is the first time a Chlamydia vaccine has been tested in the male koala and the first assessment of a mucosal vaccination route in this species. Our results suggest that vaccination of male koalas can elicit mucosal immunity and could contribute to the long-term survivability of wild populations of the koala., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Persistent organic pollutants (POPs), such as toxic lipophilic organochlorine (OC) compounds, accumulate in the blubber tissue of marine mammals. Toxicological sampling methods most frequently target only the superficial blubber layer. Vertical distribution of these contaminants through the blubber mantle may, however, not be homogenous and could reflect any dissemination of lipids and fatty acids (FAs). It is therefore critical to assess stratification patterns in a species of interest as a quality control measure for interpretation of toxicological data. Here, we analysed and compared the distribution of lipids, FAs, and OCs in the outermost and innermost blubber layer of southern hemisphere humpback whales. FA stratification was evident for short-chain (≤18) monounsaturated fatty acids (SC-MUFA), which were concentrated in the outer layer, consistent with the thermoregulatory role of this blubber layer. This stratification was, however, not reflected in OC distribution, which was similar in the inner and outer blubber layers of male humpback whales. By comparison, a noticeable gradient in total blubber lipid from the outer to the inner layer was observed in two lactating females, which coincided with higher lipid normalised contaminant levels in the inner layer. This study contains the most comprehensive assessment of humpback whale blubber stratification to date, however, further investigation of biological and ecological influencing factors is required., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Southern hemisphere humpback whales undertake the longest migrations and associated periods of fasting of any mammal. Fluctuations in lipid energy stores are known to profoundly affect the toxicokinetics of lipophilic organochlorine compound (OC) burdens. Results from blubber biopsy sampling of adult, male humpback whales at two time points of the annual migration journey revealed dramatic concentration effects for the majority of OC compounds. The observed concentration effect was, however, not linear with measured average blubber lipid loss indicating significant redistribution of OCs and hence the importance of alternate lipid depots for meeting the energetic demands of the migration journey. Applying lipophilic OC burdens as novel tracers of whole-body lipid dynamics, the observed average concentration index suggests an average individual weight loss of 13% over 4 months of the migration journey. This value is based upon lipid derived energy and is in good agreement with previous weight prediction formulas. Notably, however, these estimates may greatly underestimate individual weight loss if significant protein catabolism occurs. Biomagnification factors between migrating southern hemisphere humpback whales and their principal prey item, Antarctic krill, closely resembled those of baleen whales feeding on herbivorous zooplankton in the Arctic. This study emphasizes the importance of considering prolonged periods of food deprivation when assessing chemical risks posed to wildlife. This is of particular importance for Polar biota adapted to extremes in ecosystem productivity.

Large mysticete whales represent a unique challenge for chemical risk assessment. Few epidemiological investigations are possible due to the low incidence of adult stranding events. Similarly their often extreme life-history adaptations of prolonged migration and fasting challenge exposure assumptions. Molecular biomarkers offer the potential to complement information yielded through tissue chemical analysis, as well as providing evidence of a molecular response to chemical exposure. In this study we confirm the presence of cytochrome P450 reductase (CPR) and cytochrome P450 isoenzyme 1A1 (CYP1A1) in epidermal tissue of southern hemisphere humpback whales (Megaptera novaeangliae). The detection of CYP1A1 in the integument of the humpback whale affords the opportunity for further quantitative non-destructive investigations of enzyme activity as a function of chemical stress., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The population distribution and zoonotic potential of gastrointestinal helminths in a naturally infected population of wild rats (Rattus rattus and Rattus norvegicus) in Jamaica are described. One hundred and thirty (29.7%) of 437 rats captured in the study were infected: 104 (35%) of 297 R. rattus compared with 26 (18.6%) of 140 R. norvegicus. Nine species of gastrointestinal helminths were recovered: Raillietina sp. (0.2%), Trichuris sp. (0.2%), Rictularia sp. (0.7%), Syphacia obvelata (1.1%), Strongyloides ratti (1.4%), Hymenolepis diminuta (3.8%), Protospirura muricola (4.3%), Moniliformis moniliformis (11.2%), and Nippostrongylus brasiliensis (14.2%). In a logistic model, the single risk factor identified for both M. moniliformis and P. muricola was R. rattus, compared with R. norvegicus (OR = 8.369 and 9.714, respectively). In comparison, the risk factor predicted for infection with N. brasiliensis was the northeastern section of Jamaica (OR = 11.000) compared with western Jamaica. Rictularia sp. represents a new geographic distribution record for the Caribbean region. Hymenolepis diminuta, M. moniliformis, Raillietina sp., and Rictularia sp. are potentially zoonotic, but only human infection with H. diminuta has been previously reported in the Caribbean.

Short-beaked echidnas have an impressive ability to submerge completely into soil or sand and remain there, cryptic, for long periods. This poses questions about how they manage their respiration, cut off from a free flow of gases. We measured the gradient in oxygen partial pressure (PO2) away from the snouts of buried echidnas and oxygen consumption (VO2) in five individuals under similar conditions, in two substrates with different air-filled porosities (fa). A theoretical diffusion model indicated that diffusion alone was insufficient to account for the flux of oxygen required to meet measured rates of VO2. However, it was noticed that echidnas often showed periodic movements of the anterior part of the body, as if such movements were a deliberate effort to flush the tidal air space surrounding their nostrils. These ;flushing movements' were subsequently found to temporarily increase the levels of interstitial oxygen in the soil around the head region. Flushing movements were more frequent while VO2 was higher during the burrowing process, and also in substrate with lower fa. We conclude that oxygen supply to buried echidnas is maintained by diffusion through the soil augmented by periodic flushing movements, which ventilate the tidal airspace that surrounds the nostrils.