Table 1.
SIMPER analysis indicating the percent contribution (Ct%) of specific variables to the observed site [Rottnest Island (RI) and mainland (ML)] differences in (a) Haematology (HMT), (b) Blood chemistry (BLC), and (c) Peripheral blood cell morphology (PBCM) profiles of quokkas. For (a and b), mean with standard deviation. For (c), the number of animals (percentage of sampled animals) with 95% confidence intervals (§).
Fig 1.
Photomicrographs of reference leukocytes and red blood cells observed in blood smears of quokkas on Rottnest Island and the mainland of Western Australia.
(a) neutrophil, key features: polylobated nucleus with 3–6 lobes, coarsely clumped chromatin, primary cytoplasmic granules -azurophilic-; also present, atypical arrangement of erythrocytes: Rouleaux formation (arrowheads); (b) eosinophil, key features: polylobated nucleus with 2–3 lobes, coarsely clumped chromatin but less dense than neutrophils, prominent secondary eosinophilic cytoplasmic granules; (c) basophil, key features: intense secondary basophilic cytoplasmic granules; (d) lymphocytes, key features: darkly staining chromatin with no apparent nucleolus, cytoplasm presents as a “rim” and appears finely or coarsely granular, nuclear:cytoplasm (N:C) ratio smaller than that of other leukocytes; (e) monocyte, key features: indented to irregularly shaped nucleus, reticular chromatin, large amount of pale, grey to basophilic cytoplasm, often with vacuoles, overall size of the cell is greater than all other leukocytes; a keratocyte (i.e. poikilocyte; arrowhead) is also present; (f) erythrocytes with normal morphology (arrowheads), and with erythtocytic inclusions (e.g. Heinz body -arrow-). All photomicrographs: original magnification x1000-, staining Wright-Giemsa.
Fig 2.
Correlation (r) of white blood cell classification by the ADVIA-120® with the 200-cell manual differential method.
Eosinophils (a. r = 0.41), Basophils (b. r = 0.153), Monocytes (c. r = 0.04), Neutrophils (d. r = 0.90), and Lymphocytes (e. r = 0.93). Distances along the axes are unit-less, therefore the positions of the points in the plots are relative distances from one another rather than absolute differences read in these units.
Fig 3.
Non-metric MDS plots illustrating the structural dissimilarity in (a) haematology HMT (RI n = 96, ML n = 32), (b) blood chemistry BLC (RI n = 106, ML n = 32), and (c) peripheral blood cell morphology PBCM (RI n = 107, ML n = 34) for quokkas from Rottnest Island (open symbols) and the mainland (filled symbols) (left hand panel) or quokkas on Rottnest Island sampled at four times of the year (‘seasons’: autumn: triangles, winter: squares, spring: circles, and summer: diamonds) (right-hand panel). Key legend applies for all plots. Note that the distances along the axes are unit-less, therefore the positions of the points in the plots are relative distances from one another rather than absolute differences read in these units.
Table 2.
Summary of two-way ANOSIM (R statistic and p value) of blood analytes (see footnote) for Rottnest Island (RI) and mainland (ML) (left hand column) and Rottnest Island only (right hand column) quokkas.
Fig 4.
Linear plots for haematocrit (HMT) analytes across seasons for quokkas trapped on Rottnest Island between March and December 2011.
Autumn n = 20, Winter n = 21, Spring n = 27, Summer n = 28.
Fig 5.
Linear plots for blood chemistry (BLC) analytes across seasons for quokkas trapped on Rottnest Island between March and December 2011.
Autumn n = 26, Winter n = 25, Spring n = 26, Summer n = 29.
Fig 6.
Some erythrocytic morphologies and intraerythrocytic inclusions found in peripheral blood smears of quokkas on Rottnest Island (also found in quokkas on mainland Western Australia).
(a) echinocytes (arrow), two large cells present: neutrophil (left), eosinophil (right); (b) Heinz bodies (arrowheads), blister polychromatophilic cell (arrow), anisocytosis is also present in this plate; (c) Rouleaux formation (arrowheads), lymphocyte with a flower-like nucleus (large cell centre); (d) keratocyte (arrow), monocyte (centre) and polychromatophilic erythrocytes present; (e) Howell-Jolly body (arrow), leukocyte with fragmented nucleus (large cell centre). All images: magnification x1000, staining Wright-Giemsa.
Table 3.
SIMPER analysis indicating the percent contribution (Ct %) of specific variables to the observed time of year differences in (a) Haematology (HMT), (b) Blood chemistry (BLC), and (c) Peripheral blood cell morphology (PBCM) profiles of quokkas from Rottnest Island. For (a and b), mean and standard deviation. For (c), the number of animals (percentage of sampled animals) with 95% confidence intervals(§). Abbreviations and units as per Table 1.
Fig 7.
Box-plot of the concentration of vitamin E (mg/L) in plasma in winter for quokkas from Rottnest Island (free-ranging, n = 29) and quokkas from Perth Zoo (captive, n = 8).
Fig 8.
Microphotographs of atypical lymphocytes observed in peripheral blood smears of quokkas on Rottnest Island (a-c) atypical lymphocytes exhibiting a “flower” shape-like polylobated nucleus with condensed homogeneous chromatin; (d) atypical lymphocyte exhibiting a polylobated nucleus resembling “prototype lymphocytes”; plates a-d: original magnification x1000, stain: Wright-Giemsa.
Table 4.
Summary of mixed infections captured in both haematology and blood chemistry datasets combined, for Rottnest Island and mainland animals that tested positive (+) for eight infectious agents.
Table 5.
Correlations between three axes of a 3D non-metric Multidimensional Scaling (nMDS) model for (a) haematology (HMT) and (b) blood chemistry (BLC) analytes for Rottnest Island and mainland animals (combined sample) that were tested for eight infectious agents (IA). In this case, the three-dimensional model had less stress than the two-dimensional model. Values above the horizontal line for HMT and BLC are Spearman rank order correlation coefficients with the nMDS axes scores. Below the horizontal lines are results of multiple regression testing for relationships with site and the presence/absence of six (HMT) or five (BLC) of these IA. Colour shading indicates high to low correlations of HMT (red to blue) and BLC (red to green).
Fig 9.
Cluster analysis and patterns of 10 HMT analytes and the three nMDS axis scores derived from these values for 12 animals from the mainland (M) and 44 animals from Rottnest Island (R) (each row represents an individual).
Animals that tested positive for one of eight Infectious Agents (Salmonella spp., microfilariae, MaHV-6, Theileria spp., Cryptococcus spp., saprophyte fungi, Babesia spp., and trypanosomes) are shown as a black square. Clusters A (Rottnest Island), B (mainland), C (coinfection with Salmonella–microfilariae; green dotted line), D (minimal infections), E (coinfection with saprophytic fungi and other organisms) and F (coinfection with other organisms but not with saprophytic fungi). Salmonella–microfilariae negative animals (red dotted line).
Fig 10.
Cluster analysis and patterns of 16 BLC analytes of 47 animals from Rottnest Island (R) and 9 animals from the mainland (M) that were tested for seven infectious agents (Salmonella spp., microfilariae, MaHV-6, Theileria spp., Cryptococcus spp., saprophyte fungi and Babesia spp.
Trypanosomes were not included as there were no positive animals). Clusters A (Rottnest Island), B (mainland), C (microfilariae-positive and coinfection with MaHV-6, Theileria spp., and saprophyte fungi), and D (microfilariae-negative and coinfection with MaHV-6, Theileria spp. and saprophyte fungi). General patterns in axes 1, 2, and 3 combined showed differences in microfilariae-positive animals (blue dotted line), and microfilariae-negative animals (green dotted line).
Table 6.
Haematology and blood chemistry reference intervals for anaesthetised free-ranging quokkas sampled on the mainland of Western Australia between September 2010 and July 2011.
Intervals calculated as being less than zero were set to zero (bold). Abbreviations and units as per Table 1.
Table 7.
Haematology and blood chemistry reference intervals for anaesthetised free-ranging quokkas sampled on Rottnest Island (RI) between March and December 2011.
Intervals calculated as being less than zero were set to zero (bold). Abbreviations and units as per Table 1.