Retrospective pathological survey of pulmonary disease in free-ranging river dolphins in Brazil

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1 Thaís Carneiro Santos Rodrigues, José Luis Catão-Dias, Josué Díaz-Delgado, Júlia Luz Carvalho, Miriam Marmontel Retrospective pathological survey of pulmonary disease in free-ranging river dolphins in Brazil

2 INTRODUCTION Inia geoffrensis Dorsal fin long but low in height Unfused cervical vertebrae Dark-grey to pink Largest species of river dolphin Adult male can reach 2,55 m and 200 kg

3 INTRODUCTION Sotalia fluviatilis Dorsal fin falcate, tall but short in length Fused cervical vertebrae Light to dark grey Ventral region lighter or pinkish Smallest delphinid 1,5 m max length and weight up to 50 kg The only exclusively freshwater delphinid

4 INTRODUCTION Endemic of the Amazon and Orinoco basins Brazil, Colombia, Ecuador, Peru, Bolivia and Venezuela Endemic of the Amazon basin Brazil, Colombia, Ecuador and Peru

5 INTRODUCTION Indicators of Amazon s aquatic ecosystem quality Keep fish populations healthy and balanced

6 INTRODUCTION Limited knowledge?

7 INTRODUCTION (2014) Ameaçada

INTRODUCTION Major known threats Incidental mortality in fishing nets Population fragmentation, isolation, habitat loss *dams Environmental

8 INTRODUCTION Major known threats Incidental mortality in fishing nets Population fragmentation, isolation, habitat loss *dams Environmental pollution *mercury Inia Deliberate killing for use as fish bait for catfish (Calophysus macropterus) and to avoid (perceived) fishing competition

9 INTRODUCTION AND OBJECTIVE Underlying pre-existing diseases may contribute towards death Pneumonia and other respiratory diseases have been recognized as important causes of death in both captive and free-ranging cetaceans Survey on worldwide captive I. geoffrensis: pneumonia as the most common finding (Bonar et al., 2007) Limited information on free-ranging I. geoffrensis and S. fluviatilis Pathological survey of pulmonary disease in free-ranging Amazon river dolphin and tucuxi

10 MATERIALS AND METHODS Amazonas state

11 MATERIALS AND METHODS Dry season Wet season

12 MATERIALS AND METHODS MSDR and ASDR Riverside communities

13 MATERIALS AND METHODS Carcass collection Necropsy

14 MATERIALS AND METHODS 10% buffered formalin Histopatological process H&E Gram stain Parasitological identification (Delyamure, 1955)

RESULTS AND DISCUSSION 52 22 22 25 adults

15 RESULTS AND DISCUSSION adults 16 juveniles 6 neonates

16 RESULTS AND DISCUSSION 84,6% (44/52) lung samples Most common respiratory disease PNEUMONIA 53,8% Verminous pneumonia 25% (13/52) Bacterial pneumonia 25% (13/52) Meconium aspiration syndrome 1,9% (1/52)

17 RESULTS AND DISCUSSION Verminous Pneumonia 25% Inia geoffrensis 25% Sotalia fluviatilis Principalmente adultos e jovens de ambos os sexos e um neonato Multifocal, poorly demarcated, pale pink/yellow to dark grey, firm foci within the pulmonary parenchyma 2cm

18 RESULTS AND DISCUSSION Verminous Pneumonia Eosinophilic and suppurative to pyogranulomatous inflammatory exudate Granulomas and fibrosis 500µm Intra-alveolar, bronchiolar or bronchial adult nematodes

19 RESULTS AND DISCUSSION Verminous pneumonia 84,6% (11/13) Halocercus brasiliensis Case µm Sotalia fluviatilis Female Adult Tachypnea Unable to dive

20 RESULTS AND DISCUSSION Verminous pneumonia Halocercus brasiliensis FILO FAMILY Nematoda Pseudaliidae GENUS Halocercus spp. Most common lungworms in odontocete species worldwide Woodard et al., 1969; Dailey, 2001; Measure, 2001

21 RESULTS AND DISCUSSION Verminous Pneumonia Halocercus brasiliensis 40% of cases: probable cause of death 88% S. guianensis from southern and southeastern coasts of Brazil Guiana dolphin Sotalia guianensis Striped dolphins Stenella coeruleoalba Rosas et al., 2002; Marigo et al., 2010

22 RESULTS AND DISCUSSION Verminous Pneumonia Halocercus brasiliensis Vertical transmission Conlogue et al., 1985; Dailey et al., 1991; Moser e Rhinehart, 1993 Female neonate Sotalia fluviatilis Supports transplacental infestation

23 RESULTS AND DISCUSSION Bacterial pneumonia Suppurative bronchopneumonia Suppurative inflammatory exsudate Bronchiectasis, abundant gram-positive and gram-negative coccoid and bacillary bacteria Fibrosis, tissue remodeling

24 RESULTS AND DISCUSSION Bacterial pneumonia Case 20 Inia geoffrensis Male Adult Thorax penetrating wound Pyothorax Severe fibrinosuppurative pleuritis of the right lung 10cm 5cm

25 RESULTS AND DISCUSSION Bacterial pneumonia 500µm 20µm Case 20 Inia geoffrensis Male Adult Thorax penetrating wound Pyothorax Severe fibrinosuppurative pleuritis of the right lung Severe, diffuse, chronic fribinosuppurative pleuropneumonia Gram+ coccus

RESULTS AND DISCUSSION Bacterial pneumonia Several species of bacteria isolated from cetacean lungs Gram-negative Parsons e Jefferson, 2000; Higgins, 2000; Cfee e Lipscomb, 2009; Ridgway e Harrison,

26 RESULTS AND DISCUSSION Bacterial pneumonia Several species of bacteria isolated from cetacean lungs Gram-negative Parsons e Jefferson, 2000; Higgins, 2000; Cfee e Lipscomb, 2009; Ridgway e Harrison, 1998 Proteus sp., Morganella morganii, Alcaligenes sp., Citrobacter sp., Chryseobacterium sp., Escherichia coli, Pasteurella sp., Pseudomonas sp., Streptococcus sp. Bonar et al., 2007 Streptococcus iniae Bonar e Wagner, 2003; Bonar et al., 2007

27 RESULTS AND DISCUSSION Bacterial pneumonia Streptococcus iniae Suppurative bronchopneumonia Song et al., 2017 Golf ball disease Abscedative dermatitis and panniculitis Pyogenic septicemia Bonar e Wagner, 2003; Bonar et al., 2007

28 RESULTS AND DISCUSSION Bacterial pneumonia Streptococcus iniae Zoonotic importance in Mid-Solimões river Freitas e Rivas, 2006

RESULTS AND DISCUSSION Case 25 Meconium Aspiration Syndrome Sotalia fluviatilis Neonate Marked and diffuse pulmonary atelectasia Abundant

29 RESULTS AND DISCUSSION Case 25 Meconium Aspiration Syndrome Sotalia fluviatilis Neonate Marked and diffuse pulmonary atelectasia Abundant aspirated material Keratinized stratified epithelium with retained nuclei (pulmonary vernix caseosa) Brown granular aggregates (meconium) 200µm

30 RESULTS AND DISCUSSION Meconium Aspiration Syndrome Sotalia fluviatilis neonate Histopathological features described in humans and animals Van-Elk et al., 2007; Raju et al., 2010; Tanaka et al., 2014

first few breaths Miller, 1977; Srinivasan e Vidyasagar, 1999 Indicator of fetal

31 RESULTS AND DISCUSSION Meconium Aspiration Syndrome Aspiration of meconiumstained amniotic fluid in the airways Intrauterine gasping or during the first few breaths Miller, 1977; Srinivasan e Vidyasagar, 1999 Indicator of fetal distress Fetal anoxia or hypoxia Ojha et al., 2006; Alonso-Spilbury et al., 2005; Tayade, 2012

32 CONCLUSION High occurrence of pulmonary pathology in free-ranging I. geoffrensis and S. fluviatilis in the Mid-Solimões Verminous pneumonia Bacterial pneumonia First report of Halocercus brasiliensis infection in I. geoffrensis First report of Meconium Aspiration Syndrome in S. fluviatilis Understanding the impact of lung disease in these riverine species Future medical and conservation efforts on these Amazonian dolphins

Universitat Autonoma de Barcelona (UAB). Facultad de Veterinaria, Servicio de Ecopatologia de Fauna Salvaje. Barcelona, España. 6.

Universitat Autonoma de Barcelona (UAB). Facultad de Veterinaria, Servicio de Ecopatologia de Fauna Salvaje. Barcelona, España. 6. PARASITES OF SOTALIA GUIANENSIS IN THE SOUTH AND SOUTHEASTERN COASTS OF BRAZIL Marigo, J. 1,2, Ruoppolo, V. 1,3 ; Rosas, F.C.W. 4, Andrade, A.L.V. 5, Oliveira, M.R. 6, Dias, R.A. 7 and Catão-Dias, J.L.