Amanda Duarte Barbosa

Bovine mastitis remains one of the most widespread and economically significant diseases affecting the dairy sector in Pakistan. Despite being a leading global milk producer, Pakistan’s dairy industry faces persistent challenges in controlling mastitis, particularly among smallholder farmers with limited access to veterinary care and diagnostic tools. Antibiotics are frequently used to manage mastitis, often without veterinary oversight, contributing to the growing concern of antimicrobial resistance (AMR). The widespread presence of
Staphylococcus aureus
, a biofilm-forming pathogen, further complicates treatment and accelerates resistance development. Pakistan has initiated efforts to address AMR through national action plans. However, gaps remain in surveillance, responsible antibiotic use, and access to affordable alternatives. This review examines the current landscape of bovine mastitis in Pakistan, including its epidemiology, economic impact, therapeutic practices, and AMR trends. It also outlines practical, low-cost recommendations for improving mastitis management and reducing inappropriate use of antibiotics, particularly in rural settings. Addressing these interconnected challenges through locally adapted, sustainable approaches is essential for improving animal health, milk safety, and long-term productivity in Pakistan’s dairy sector, whilst addressing the global challenge of AMR.

Background
The emergence of Ehrlichia canis in northern Australia in 2020 has reshaped the landscape of tick-borne diseases in dogs, particularly in rural and remote communities where the brown dog tick (Rhipicephalus linnaei) is endemic. Despite the rapid spread of ehrlichiosis and reported impacts on dog health, its prevalence remains poorly understood. This study aims to provide baseline data on the epidemiology of E. canis in Australia by determining its seroprevalence in dogs from a remote Northern Territory community.

Methods
In a cross-sectional study, an enzyme-linked immunosorbent assay (ELISA) assessed the point seroprevalence of E. canis in community dogs. A door-to-door census was undertaken to quantify resident dogs and obtain information on signalment and clinical signs. Canine serum samples were evaluated for seroreactivity to E. canis at the state reference laboratory.

Results
Of the 48 dogs present in the community, 44 (91.2%) were included in the serosurvey. ELISA testing found a point seroprevalence of 52.3% (95% CI: 36.7% to 67.5%) for E. canis. Seropositive dogs were mostly asymptomatic and had similar body condition scores to seronegative dogs.

Conclusions
This study documents a high point seroprevalence of E. canis in dogs from a remote Australian community. There was no significant correlation between serostatus and gender, body condition or the presence of clinical signs. These results underscore the need for further research to understand the clinical significance of seropositivity in asymptomatic dogs and highlight the need for a locally validated diagnostic test to support field-based surveillance and management of ehrlichiosis in Australia.

Tick bites and tick-related diseases are on the rise. Diagnostic tests that identify well-characterised tick-borne pathogens (TBPs) possess limited capacity to address the causation of symptoms associated with poorly characterised tick-related illnesses, such as debilitating symptom complexes attributed to ticks (DSCATT) in Australia. Identification of local signals in tick-bitten skin that can be detected systemically in blood would have both clinical (diagnostic or prognostic) and research (mechanistic insight) utility, as a blood sample is more readily obtainable than tissue biopsies. We hypothesised that blood samples may reveal signals which reflect relevant local (tissue) events and that the time course of these signals may align with local pathophysiology. As a first step towards testing this hypothesis, we compared molecular signatures in skin biopsies taken from the tick-bite location of human participants, as published in our previous study, together with peripheral blood signatures obtained concurrently. This approach captures differentially expressed molecules across multiple omics datasets derived from peripheral blood (including cellular and cell-free transcriptomics, proteomics, metabolomics, and DNA methylation), and skin biopsies (spatial transcriptomics). Our original data revealed that extracellular matrix organisation and platelet degranulation pathways were upregulated in the skin within 72 h of a tick bite. The same signals appeared in blood, where they then remained elevated for three months, displaying longitudinally consistent alterations of biological functions. Despite the limited sample size, these data represent proof-of-concept that molecular events in the skin following a tick bite can be detectable systemically. This underscores the potential value of blood samples, akin to a liquid biopsy, to capture biomarkers reflecting local tissue processes.

The collection of baseline health data is an essential component of an endangered species conservation program. As for many wildlife species, there are minimal health data available for wild populations of the endangered Carnaby’s cockatoo (Zanda latirostris). In this study, 426 wild Carnaby’s cockatoo nestlings were sampled from nine breeding sites throughout the range of the species over an 11-year period. In addition to a physical examination, samples were collected to test for hematologic and biochemical parameters, psittacine beak and feather disease virus (BFDV), avian polyomavirus (APV), psittacine adenovirus, psittacine herpesvirus, Chlamydia, disease serology and endoparasites. Environmental sampling was performed to screen for BFDV and APV in nest hollows. Descriptive health data are presented for nestlings of this species, with BFDV, APV and Chlamydia infections reported. Reference intervals for hematologic and biochemical parameters are presented in three age groups, and factors affecting blood analytes and body condition index are discussed. This longitudinal dataset provides insights into health parameters for Carnaby’s cockatoo nestlings and a reference for future monitoring of breeding populations.

Marsupials, inhabiting diverse ecosystems, including urban and peri-urban regions in Australasia and the Americas, intersect with human activities, leading to zoonotic spill-over and anthroponotic spill-back of pathogens, including Cryptosporidium and Giardia. This review assesses the current knowledge on the diversity of Cryptosporidium and Giardia species in marsupials, focusing on the potential zoonotic risks. Cryptosporidium fayeri and C. macropodum are the dominant species in marsupials, while in possums, the host-specific possum genotype dominates. Of these three species/genotypes, only C. fayeri has been identified in two humans and the zoonotic risk is considered low. Generally, oocyst shedding in marsupials is low, further supporting a low transmission risk. However, there is some evidence of spill-back of C. hominis into kangaroo populations, which requires continued monitoring. Although C. hominis does not appear to be established in small marsupials like possums, comprehensive screening and analysis are essential for a better understanding of the prevalence and potential establishment of zoonotic Cryptosporidium species in small marsupials. Both host-specific and zoonotic Giardia species have been identified in marsupials. The dominance of zoonotic G. duodenalis assemblages A and B in marsupials may result from spill-back from livestock and humans and it is not yet understood if these are transient or established infections. Future studies using multilocus typing tools and whole-genome sequencing are required for a better understanding of the zoonotic risk from Giardia infections in marsupials. Moreover, much more extensive screening of a wider range of marsupial species, particularly in peri-urban areas, is required to provide a clearer understanding of the zoonotic risk of Cryptosporidium and Giardia in marsupials.

Rodents represent the single largest group within mammals and host a diverse array of zoonotic pathogens. Urbanisation impacts wild mammals, including rodents, leading to habitat loss but also providing new resources. Urban-adapted (synanthropic) rodents, such as the brown rat (R. norvegicus), black rat (R. rattus), and house mouse (Mus musculus), have long successfully adapted to living close to humans and are known carriers of zoonotic pathogens. Two important enteric, zoonotic protozoan parasites, carried by rodents, include Cryptosporidium and Giardia. Their environmental stages (oocysts/cysts), released in faeces, can contaminate surface and wastewaters, are resistant to common drinking water disinfectants and can cause water-borne related gastritis outbreaks. At least 48 species of Cryptosporidium have been described, with C. hominis and C. parvum responsible for the majority of human infections, while Giardia duodenalis assemblages A and B are the main human-infectious assemblages. Molecular characterisation is crucial to assess the public health risk linked to rodent-related water contamination due to morphological overlap between species. This review explores the global molecular diversity of these parasites in rodents, with a focus on evaluating the zoonotic risk from contamination of water and wasterwater with Cryptosporidium and Giardia oocysts/cysts from synanthropic rodents. Analysis indicates that while zoonotic Cryptosporidium and Giardia are prevalent in farmed and pet rodents, host-specific Cryptosporidium and Giardia species dominate in urban adapted rodents, and therefore the risks posed by these rodents in the transmission of zoonotic Cryptosporidium and Giardia are relatively low. Many knowledge gaps remain however, and therefore understanding the intricate dynamics of these parasites in rodent populations is essential for managing their impact on human health and water quality. This knowledge can inform strategies to reduce disease transmission and ensure safe drinking water in urban and peri‑urban areas.

Rabbits are highly abundant in many countries and can serve as reservoirs of diseases for a diversity of pathogens including the enteric protozoan parasites, Cryptosporidium and Giardia. Both parasites shed environmentally robust environmental stages (oo/cysts) and have been responsible for numerous waterborne outbreaks of diseases. Cryptosporidium hominis and C. parvum are responsible for most infections in humans, while Giardia duodenalis assemblages A and B, cause most human cases of giardiasis. Cryptosporidium cuniculus, the dominant species infecting rabbits, is the only spceies other than C. hominis and C. parvum to have caused a waterborne outbreak of gastritis, which occurred in the United Kingdom in 2008. This review examines the prevalence of Cryptosporidium and Giardia species in rabbits to better understand the public health risks of contamination of water sources with Cryptosporidium and Giardia oo/cysts from rabbits. Despite the abundance of C. cuniculus in rabbits, reports in humans are relatively rare, with the exception of the United Kingdom and New Zealand, and reports of C. cuniculus in humans from the United Kingdom have declined substantially since the 2008 outbreak. Subtyping of C. cuniculus has supported the potential for zoonotic transmission. Relatively few studies have been conducted on Giardia, but assemblage B dominates. However, improved typing methods are required to better understand the transmission dynamics of Giardia assemblages in rabbits. Similarly, it is not well understood if pet rabbits or contaminated water are the main source of C. cuniculus infections in humans. Well-planned studies using high-resolution typing tools are required to understand the transmission dynamics better and quantify the public health risk of Cryptosporidium and Giardia from rabbits.

Bovine anaemia caused by Theileria orientalis group (BATOG) causes significant production and economic losses in Australia's cattle industry. The pathogenic T. orientalis genotypes reported in Australian cattle are type 1 (Chitose) and type 2 (Ikeda). The present study aimed to determine the prevalence and distribution of T. orientalis genotypes in adult lactating cows in Western Australia (WA) dairy herds. A total of 100 whole blood samples from lactating cows from 10 farms were obtained and screened for T. orientalis using polymerase chain reaction (PCR). Sanger sequencing was subsequently used to characterise T. orientalis genotypes isolated from positive samples. A total of thirteen cows (13%; 95% CI: 7.1-21.2%) were positive for T. orientalis, and six out of ten farms (60%; 95% CI: 26.2-87.8%) housed at least one T. orientalis-positive cow. The distribution of T. orientalis was found to be wide and dense in the South west region of WA and the southern coast of WA. The predominant T. orientalis genotype identified was Ikeda (n = 11, 11%; 95% CI: 5.6-18.8%), while the Buffeli genotype was identified in WA for the first time, albeit at a low prevalence (n = 1, 1%; 95% CI: 0.0-5.4%). This study has provided useful epidemiological evidence on the prevalence and distribution of T. orientalis in adult lactating dairy cows in WA dairy farms, and on the importance of conducting widespread surveillance programs for the understanding of BATOG in WA.

Bats are known to harbour various pathogens and are increasingly recognised as potential reservoirs for zoonotic diseases. This paper reviews the genetic diversity and zoonotic potential of Cryptosporidium and Giardia in bats. The risk of zoonotic transmission of Cryptosporidium from bats to humans appears low, with bat-specific Cryptosporidium genotypes accounting for 91.5% of Cryptosporidium-positive samples genotyped from bats worldwide, and C. parvum and C. hominis accounting for 3.4% each of typed positives, respectively. To date, there have only been sporadic detections of Giardia in bats, with no genetic characterisation of the parasite to species or assemblage level. Therefore, the role bats play as reservoirs of zoonotic Giardia spp. is unknown. To mitigate potential risks of zoonotic transmission and their public health implications, comprehensive research on Cryptosporidium and Giardia in bats is imperative. Future studies should encompass additional locations across the globe and a broader spectrum of bat species, with a focus on those adapted to urban environments.

Cryptosporidium and Giardia are important waterborne protozoan parasites that are resistant to disinfectants commonly used for drinking water. Wild birds, especially wild migratory birds, are often implicated in the contamination of source and wastewater with zoonotic diseases, due to their abundance near water and in urban areas and their ability to spread enteric pathogens over long distances. This review summarises the diversity of Cryptosporidium and Giardia in birds, with a focus on zoonotic species, particularly in wild and migratory birds, which is critical for understanding zoonotic risks. The analysis revealed that both avian-adapted and zoonotic Cryptosporidium species have been identified in birds but that avian-adapted Cryptosporidium species dominate in wild migratory birds. Few studies have examined Giardia species and assemblages in birds, but the non-zoonotic Giardia psittaci and Giardia ardeae are the most commonly reported species. The identification of zoonotic Cryptosporidium and Giardia in birds, particularly C. parvum and G. duodenalis assemblages A and B in wild migratory birds, is likely due to mechanical carriage or spillback from birds co-grazing pastures contaminated with C. parvum from livestock. Therefore, the role of wild migratory birds in the transmission of zoonotic Cryptosporidium and Giardia to source water is likely overestimated. To address knowledge gaps, it is important to conduct more extensive studies on the prevalence of Cryptosporidium and Giardia in a broader range of migratory wild birds. There is also a need to investigate the extent to which zoonotic infections with C. hominis/C. parvum and G. duodenalis assemblages A and B are mechanical and/or transient, and to assess the load and viability of zoonotic oo/cysts shed in avian faeces. Understanding the contribution of birds to zoonoses is essential for effective disease surveillance, prevention, and control.